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Home , Ets variant 5, Glyoxylate reductase, Glypican 5, GNA14, GNAL, Interleukin 2

Supplemental Data

Article's title: Purinergic by the β2-adrenergic receptor increases intracellular Ca2+ in non-excitable cells

Author's names: Wayne Stallaert, Emma T van der Westhuizen, Anne-Marie Schönegge, Bianca Plouffe, Mireille Hogue, Viktoria Lukashova, Asuka Inoue, Satoru Ishida, Junken Aoki, Christian Le Gouill & Michel Bouvier

Journal title: Molecular Pharmacology

1 Supplemental Table 1 – Primer sequences and restriction combinations to screen for mutants.

Target Primer 1 Primer 2 Enzyme

GNAS 5’-TCAACGGTAGGATGCTGTGG-3’ 5’-CTACAAGAAGGGAGGCCGTG-3’ Hap II

GNAL #1 5’-AAGCACGTTTGCCATTGTCC-3’ 5’-CTTCAGGTTATCCGCCCTCC-3’ Hae III

GNAL #2 5’-ACTGTCACCAAAGCCTCCAG-3’ 5’-TACTGCTTGAGGTGCATCCG-3’ Hap II

GNAL #3 5’-ACACTAAACATAGAGTGGGTGC-3’ 5’-TGAACAAAACTTTCTGGTTGTCAG-3’ Pvu II

2 Supplemental Table 2: List of found to be significantly up or down regulated in the ΔGs cells (clone 1) with a minimum of 2 fold change (<-2 or >2) vs the parental cells. Significance was established using unpaired one-way ANOVA between-samples, using the transcriptome analysis console from Affymetrix (p < 0.05 was considered statistically significant). The false discovery rate (FDR) adjustment has been calculated for each and is indicated.

Fold Change FDR Gene Description (linear) p-value Symbol (parental vs. ∆Gs) (parental vs ∆Gs) SHISA2 shisa family member 2 11.41 0.110671 PADI3 peptidyl arginine deiminase, type III -2.03 0.110671 ANKRD1 repeat domain 1 (cardiac muscle) -16.70 0.110671 CNTFR ciliary neurotrophic factor receptor 2.82 0.110671 CD44 CD44 molecule (Indian blood group) -3.47 0.112203 MPPED2 metallophosphoesterase domain containing 2 2.11 0.112203 TCF24 24 5.14 0.112203 KCTD12 potassium channel tetramerization domain containing 12 36.44 0.112203 EFNB2 ephrin-B2 6.82 0.112203 RPS6KA3 ribosomal S6 kinase, 90kDa, polypeptide 3 -2.14 0.112203 GDF11 growth differentiation factor 11 2.09 0.112203 ERVMER34-1 endogenous retrovirus group MER34, member 1 4.41 0.112203 GJB2 gap junction protein beta 2 28.92 0.112203 MRPL50 mitochondrial ribosomal protein L50 2.46 0.112203 OSMR receptor -2.81 0.112203 EPHA7 EPH receptor A7 12.08 0.112203 GPC5 glypican 5 2.29 0.112203 ODF2 outer dense fiber of sperm tails 2 3.02 0.112203 LYST lysosomal trafficking regulator -2.05 0.112203 GREM1 gremlin 1, DAN family BMP antagonist -8.63 0.112203 SRPX2 sushi-repeat containing protein, X-linked 2 -7.93 0.112203 MMP2 matrix metallopeptidase 2 -4.43 0.112203 EMP3 epithelial 3 -2.41 0.112203 SERPINF1 peptidase inhibitor, clade F (alpha-2 antiplasmin, pigment epithelium derived factor), member 1 4.50 0.112203 SLC6A9 solute carrier family 6 (neurotransmitter transporter, glycine), member 9 2.01 0.112203 SP100 SP100 nuclear -2.58 0.112203 FAXDC2 fatty acid hydroxylase domain containing 2 -2.01 0.112203 CPS1 carbamoyl-phosphate synthase 1 -3.74 0.112203 NR6A1 subfamily 6, group A, member 1 2.92 0.112203 NME5 NME/NM23 family member 5 5.70 0.112203 NALCN sodium leak channel, non selective -4.25 0.112203 IL2RG 2 receptor, gamma -6.77 0.112203 ISOC1 isochorismatase domain containing 1 2.04 0.112438 MLLT11 myeloid/lymphoid or mixed-lineage leukemia; translocated to, 11 -3.38 0.112438 TMEM154 transmembrane protein 154 -2.47 0.113586 GGT1 gamma-glutamyltransferase 1 -4.26 0.113586 STX3 syntaxin 3 -2.01 0.113586 MXRA8 matrix-remodelling associated 8 3.07 0.113586 ALPPL2 alkaline phosphatase, placental like 2 2.41 0.114411 TLR6 toll-like receptor 6 -2.53 0.114411 TRIB2 tribbles pseudokinase 2 -2.50 0.114411 DNAJC3 DnaJ (Hsp40) homolog, subfamily C, member 3 -2.04 0.114411 PAX7 paired box 7 -2.05 0.114411 HHIP hedgehog interacting protein 2.08 0.114411 BMP2 bone morphogenetic protein 2 -5.67 0.114411 ROR2 receptor kinase-like orphan receptor 2 5.05 0.114411 F10 coagulation factor X -2.02 0.114411 PARP14 poly(ADP-ribose) polymerase family member 14 -8.88 0.114411 PDGFD platelet derived D -2.12 0.114411 NFKBIA nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha -3.53 0.114411 KNOP1 lysine-rich nucleolar protein 1 2.26 0.114411 H1F0 H1 histone family, member 0 -2.67 0.114771 TINAGL1 tubulointerstitial nephritis antigen-like 1 -2.86 0.115278 RIT1 Ras-like without CAAX 1 -2.19 0.115278 RIBC2 RIB43A domain with coiled-coils 2 2.08 0.115927 GBP2 guanylate binding protein 2, -inducible -2.20 0.115927 TOR2A torsin family 2, member A 3.30 0.115927 CA2 carbonic anhydrase II 8.75 0.115927 IQGAP2 IQ motif containing GTPase 2.20 0.115992 WDR34 WD repeat domain 34 3.83 0.115992 SULT1C4 sulfotransferase family 1C member 4 10.70 0.115992 FHL2 four and a half LIM domains 2 -2.29 0.115992 GABRA3 gamma-aminobutyric acid (GABA) A receptor, alpha 3 -5.18 0.115992 ABCC2 ATP binding cassette subfamily C member 2 -2.25 0.115992 HOXB-AS3 HOXB cluster antisense RNA 3 20.44 0.119418 CITED1 Cbp/p300-interacting transactivator, with Glu/Asp rich carboxy-terminal domain, 1 4.71 0.121838 TNFRSF9 receptor superfamily, member 9 -16.22 0.122868 FILIP1L A interacting protein 1-like -2.09 0.122868 AMBN ameloblastin 5.54 0.122868 GGT2 gamma-glutamyltransferase 2 -2.08 0.125061 OLFM1 olfactomedin 1 -15.15 0.125061 VRK1 vaccinia related kinase 1 2.43 0.125061 YPEL3 yippee like 3 -2.03 0.125061 CNKSR1 connector enhancer of kinase suppressor of Ras 1 3.26 0.125061 TBC1D13 TBC1 domain family, member 13 2.95 0.125061 GPRC5C -coupled receptor, class C, group 5, member C -2.09 0.125061 ZBTB7B and BTB domain containing 7B -2.15 0.125061 GREM1 gremlin 1, DAN family BMP antagonist [Source:HGNC Symbol;Acc:HGNC:2001] -5.89 0.125061 EDIL3 EGF-like repeats and discoidin I-like domains 3 -2.76 0.125061 Supplemental Table 2: continue

MYO5B VB -2.24 0.125061 TCIRG1 T-cell, immune regulator 1, ATPase, H+ transporting, lysosomal V0 subunit A3 -2.03 0.125061 LINC00649 long intergenic non-protein coding RNA 649 7.92 0.125061 MXD1 MAX dimerization protein 1 -2.65 0.125061 SMAD7 SMAD family member 7 -2.05 0.125061 FAM111A family with sequence similarity 111, member A -3.67 0.125061 SPINT1 peptidase inhibitor, Kunitz type 1 -3.30 0.125061 MYL9 9 7.77 0.125061 LGALS1 lectin, galactoside-binding, soluble, 1 -4.66 0.125061 TAPBP TAP binding protein (tapasin) -2.07 0.129017 LRRN1 leucine rich repeat neuronal 1 -3.64 0.129017 MOB3B MOB kinase activator 3B 3.46 0.129017 TGM2 transglutaminase 2 -2.11 0.129017 SLC27A4 solute carrier family 27 (fatty acid transporter), member 4 2.63 0.129017 INA neuronal protein, alpha 6.31 0.129017 CXCL10 (C-X-C motif) ligand 10 -2.64 0.129017 C1QTNF1 C1q and tumor necrosis factor related protein 1 -3.53 0.129017 DOLK dolichol kinase 2.54 0.129017 TRUB2 TruB pseudouridine (psi) synthase family member 2 2.66 0.129017 DDR1; discoidin domain receptor tyrosine kinase 1; -2.07 0.129017 MIR4640 microRNA 4640 FAM174B family with sequence similarity 174, member B -2.20 0.129179 RGS9 regulator of G-protein signaling 9 -3.45 0.129920 KDM5B lysine (K)-specific demethylase 5B -2.04 0.130577 ZNF711 zinc finger protein 711 2.40 0.131949 HSPB8 heat shock 22kDa protein 8 -2.52 0.133236 MFAP2 microfibrillar associated protein 2 4.33 0.133506 NFKB2 nuclear factor of kappa light polypeptide gene enhancer in B-cells 2 (p49/p100) -3.33 0.133506 NDUFAF2 NADH dehydrogenase (ubiquinone) complex I, assembly factor 2 2.09 0.133506 TFAP2A transcription factor AP-2 alpha (activating enhancer binding protein 2 alpha) -2.02 0.133506 ADAM28 ADAM metallopeptidase domain 28 -2.55 0.133506 FAM27E3 family with sequence similarity 27, member E3 5.35 0.133506 CAV1 caveolin 1 -2.69 0.133506 COQ4 coenzyme Q4 3.77 0.133506 ACSM3 acyl-CoA synthetase medium-chain family member 3 2.08 0.135266 EFNA1 ephrin-A1 -2.24 0.135266 PCYT1B phosphate cytidylyltransferase 1, choline, beta -2.08 0.136233 KIAA1217 KIAA1217 -2.36 0.136233 DTX4 deltex 4, E3 ubiquitin 2.29 0.136366 HBP1 HMG-box transcription factor 1 -2.06 0.137726 POU6F2 POU class 6 2 3.42 0.138586 ACTBL2 , beta-like 2 -6.53 0.138788 NEK6 NIMA-related kinase 6 2.27 0.139162 RPP25L ribonuclease P/MRP 25kDa subunit-like 2.72 0.140728 FEZF1 FEZ family zinc finger 1 -3.16 0.140857 SPTLC3 serine palmitoyltransferase, long chain base subunit 3 -8.29 0.140857 PHF14 PHD finger protein 14 2.33 0.141430 SV2A synaptic vesicle glycoprotein 2A -2.33 0.141430 PRDM6 PR domain containing 6 3.28 0.141430 SDC4 syndecan 4 -3.34 0.141482 RUNX3 runt-related transcription factor 3 2.50 0.141482 NEFL , light polypeptide 2.83 0.142040 DDT D-dopachrome tautomerase 2.10 0.142188 ACADVL acyl-CoA dehydrogenase, very long chain -2.01 0.143238 TNFRSF12A tumor necrosis factor receptor superfamily, member 12A -2.43 0.143238 KRT17 17, type I -2.56 0.143625 SERPINB8 serpin peptidase inhibitor, clade B (), member 8 -4.11 0.146230 TNFAIP3 tumor necrosis factor, alpha-induced protein 3 -3.62 0.146230 METTL7A methyltransferase like 7A 2.09 0.146230 IER3 immediate early response 3 -7.50 0.149465 URM1 ubiquitin related modifier 1 3.77 0.149465 CASP4 caspase 4 -10.88 0.149833 IL13RA2 receptor, alpha 2 -2.55 0.149863 FLT1 fms-related tyrosine kinase 1 2.48 0.150351 ETV5 -5.05 0.151176 PAMR1 peptidase domain containing associated with muscle regeneration 1 -3.90 0.153221 PLTP phospholipid transfer protein 7.87 0.153221 NUAK2 NUAK family, SNF1-like kinase, 2 -2.95 0.153221 LAMP1 lysosomal-associated membrane protein 1 -2.23 0.153221 CCL2 chemokine (C-C motif) ligand 2 -12.25 0.153262 TMCO3 transmembrane and coiled-coil domains 3 -2.13 0.154608 NCKAP1L NCK-associated protein 1-like -2.74 0.154608 SLAIN1 SLAIN motif family member 1 3.82 0.155068 GLE1 GLE1 RNA export mediator 4.51 0.155068 EYA1 EYA transcriptional coactivator and phosphatase 1 -2.62 0.155068 MAL2 mal, T-cell differentiation protein 2 (gene/pseudogene) -3.04 0.155068 SPON1 spondin 1, extracellular matrix protein 2.27 0.155068 MAPKAP1 mitogen-activated associated protein 1 2.43 0.155068 YDJC YdjC homolog (bacterial) 2.01 0.155068 TP53INP1 tumor protein inducible nuclear protein 1 -2.46 0.155068 GPR160 G protein-coupled receptor 160 -2.01 0.155068 RINL Ras and interactor like -3.72 0.155068 EID2B EP300 interacting inhibitor of differentiation 2B 2.02 0.155068 SP140L SP140 nuclear body protein-like -4.04 0.155068 SVIL supervillin -2.24 0.155068 GCOM1; MYZAP; POLR2M GRINL1A complex locus 1; myocardial zonula adherens protein; polymerase (RNA) II (DNA directed) polypeptide M -2.01 0.155124 FABP5 fatty acid binding protein 5 (-associated) 2.22 0.155124 DUSP9 dual specificity phosphatase 9 2.03 0.155124 TENM1 teneurin transmembrane protein 1 4.35 0.155124 PPP1R15A 1, regulatory subunit 15A -2.55 0.155124 ZER1 zyg-11 related, cell cycle regulator 2.64 0.155124 Supplemental Table 2: continue

GUCY1A3 1, soluble, alpha 3 7.58 0.155124 RALGPS1 Ral GEF with PH domain and SH3 binding motif 1 2.01 0.155144 ATF3 activating transcription factor 3 -3.08 0.155596 SYT11 synaptotagmin XI -4.12 0.155596 NOV nephroblastoma overexpressed -5.79 0.155773 GALNT3 polypeptide N-acetylgalactosaminyltransferase 3 -2.73 0.155773 PRKAR2A protein kinase, cAMP-dependent, regulatory, type II, alpha 2.03 0.155773 RELB v- avian reticuloendotheliosis viral oncogene homolog B -3.11 0.155773 CD83 CD83 molecule -2.24 0.155773 GPNMB glycoprotein (transmembrane) nmb -4.50 0.155773 RAC2 ras-related C3 botulinum toxin substrate 2 (rho family, small GTP binding protein Rac2) -3.67 0.155824 MYD88 myeloid differentiation primary response 88 -7.29 0.155824 SPATA16 associated 16 16.93 0.155959 IQCK IQ motif containing K 2.09 0.156641 MYOF myoferlin -3.62 0.156641 FAM129A family with sequence similarity 129, member A -3.39 0.156641 REXO1L4P REX1, RNA exonuclease 1 homolog-like 4, pseudogene -2.33 0.156641 RNF122 ring finger protein 122 2.78 0.157160 TMEM129 transmembrane protein 129, E3 ubiquitin protein ligase 2.71 0.157433 OPTN optineurin -3.04 0.157570 GABRE; MIR224; MIR452 gamma-aminobutyric acid (GABA) A receptor, epsilon; microRNA 224; microRNA 452 -2.90 0.157570 GREM1 gremlin 1, DAN family BMP antagonist [Source:HGNC Symbol;Acc:HGNC:2001] -10.02 0.157570 CD109 CD109 molecule -3.07 0.157570 LOXL4 lysyl oxidase-like 4 -2.23 0.157570 CYBRD1 b reductase 1 -2.28 0.157570 ZBED8 zinc finger, BED-type containing 8 2.65 0.157570 DUSP5 dual specificity phosphatase 5 -2.15 0.157570 FLVCR2 subgroup C cellular receptor family, member 2 2.03 0.157570 CPA4 carboxypeptidase A4 -4.72 0.157570 CAPN6 calpain 6 -4.49 0.157845 TFPI2 tissue factor pathway inhibitor 2 -3.61 0.157845 QPCT glutaminyl-peptide cyclotransferase -2.51 0.158768 TRPC6 transient receptor potential cation channel, subfamily C, member 6 -2.10 0.158782 AIFM2 -inducing factor, mitochondrion-associated, 2 -2.07 0.158984 DERL3 derlin 3 3.09 0.160890 PSAT1 phosphoserine aminotransferase 1 2.38 0.160890 CNN1 calponin 1, basic, smooth muscle -2.60 0.161490 MALSU1 mitochondrial assembly of ribosomal large subunit 1 2.24 0.162899 LPAR5 lysophosphatidic acid receptor 5 -2.32 0.162957 GPX3 glutathione peroxidase 3 -2.67 0.162957 RGS7BP regulator of G-protein signaling 7 binding protein 2.59 0.162957 IRX4 iroquois homeobox 4 -2.35 0.162957 DACH1 dachshund family transcription factor 1 3.85 0.162957 JUNB jun B proto-oncogene -3.22 0.162957 DHRS2 dehydrogenase/reductase (SDR family) member 2 -3.84 0.162999 S100A3 S100 calcium binding protein A3 -4.88 0.162999 LIN7A lin-7 homolog A (C. elegans) 2.01 0.164273 DCBLD2 discoidin, CUB and LCCL domain containing 2 -2.12 0.164354 PTDSS2 phosphatidylserine synthase 2 2.11 0.165015 PARD6G par-6 family cell polarity regulator gamma 2.65 0.165351 TXLNG taxilin gamma 2.14 0.165737 CDC42EP3 CDC42 effector protein (Rho GTPase binding) 3 -2.11 0.167513 SPRY4 sprouty RTK signaling antagonist 4 -3.05 0.167513 IRS2 insulin receptor substrate 2 2.49 0.167513 SYTL5 synaptotagmin-like 5 -6.40 0.167769 PLCXD1 phosphatidylinositol-specific , X domain c ontaining 1 2.77 0.168301 GRHPR glyoxylate reductase/hydroxypyruvate reductase 2.07 0.168301 MAP2 associated protein 2 -2.66 0.168423 ABHD13 abhydrolase domain containing 13 -2.21 0.168423 TSPAN18 tetraspanin 18 2.19 0.168423 MVP; PAGR1 major vault protein; PAXIP1 associated glutamate-rich protein 1 -4.83 0.168423 KCNH2 potassium channel, voltage gated eag related subfamily H, member 2 -3.66 0.168423 C12orf56 12 open reading frame 56 5.71 0.168423 RCAN2 regulator of 2 10.42 0.168423 PCDH9 protocadherin 9 3.51 0.168696 SFN stratifin -2.68 0.168775 RNASE1 ribonuclease, RNase A family, 1 (pancreatic) -2.02 0.169077 HLA-DMA major histocompatibility complex, class II, DM alpha -2.95 0.169077 NR2F1 nuclear receptor subfamily 2, group F, member 1 2.14 0.169077 TMEM80 transmembrane protein 80 2.27 0.169077 selenoprotein M; selenoprotein M [Source:EntrezGene; Acc:140606]; Salzman2013 ANNOTATED, CDS, coding, OVCODE, OVERLAPTX, OVEXON, UTR3 best transcript NM_080430; Salzman2013 ANNOTATED, CDS, coding, OVCODE, SELM -2.36 0.169077 OVERLAPTX, OVEXON, UTR3, UTR5 best transcript NM_080430; Transcript Identified by AceView, Gene ID(s) 140606 HLA-DRB1 major histocompatibility complex, class II, DR beta 1 -2.47 0.169077 S100A16 S100 calcium binding protein A16 -6.34 0.169077 SEL1L3 sel-1 suppressor of lin-12-like 3 (C. elegans) -2.79 0.169077 SLC16A4 solute carrier family 16, member 4 -2.14 0.170338 CD68 CD68 molecule -2.74 0.170338 RAB26 RAB26, member RAS oncogene family 2.56 0.170338 HOXA4 homeobox A4 2.50 0.170400 HLA-DPA1 major histocompatibility complex, class II, DP alpha 1 -4.10 0.170400 PRKCB protein kinase C, beta 2.86 0.170400 HOXC8 homeobox C8 2.49 0.170611 TCEAL3 transcription elongation factor A (SII)-like 3 -2.89 0.170793 GPC3 glypican 3 2.22 0.171328 COL4A1 collagen, type IV, alpha 1 -2.11 0.172224 CD163L1 CD163 molecule-like 1 -8.28 0.172224 GBP3 guanylate binding protein 3 -2.11 0.172739 PNMAL1 paraneoplastic Ma antigen family-like 1 -2.13 0.172779 DSEL dermatan sulfate epimerase-like -5.03 0.172930 Supplemental Table 2: continue

C11orf70 open reading frame 70 2.09 0.173044 CDK9 -dependent kinase 9 2.07 0.173832 GLIPR1 GLI pathogenesis-related 1 -4.11 0.173832 KRT222 keratin 222, type II -2.40 0.174048 PUDP pseudouridine 5-phosphatase 4.97 0.174048 SLC16A7 solute carrier family 16 (monocarboxylate transporter), member 7 -3.40 0.174229 SPANXB1 SPANX family, member B1 -2.60 0.174229 ALS2CR11 amyotrophic lateral sclerosis 2 chromosome region candidate 11 2.60 0.175651 CCDC102B coiled-coil domain containing 102B -2.32 0.175738 GUCY1B3 guanylate cyclase 1, soluble, beta 3 2.96 0.175976 TNFRSF11B tumor necrosis factor receptor superfamily, member 11b -2.20 0.175976 PLCXD1 phosphatidylinositol-specific phospholipase C, X domain containing 1 2.72 0.175976 TRPV3 transient receptor potential cation channel, subfamily V, member 3 -4.30 0.175976 AIMP2 aminoacyl tRNA synthetase complex-interacting multifunctional protein 2 2.02 0.175976 NPTX2 neuronal pentraxin II 2.66 0.175976 SARM1 sterile alpha and TIR motif containing 1 2.48 0.176147 LOXL1 lysyl oxidase-like 1 -2.22 0.178033 DPM2 dolichyl-phosphate mannosyltransferase polypeptide 2, regulatory subunit 2.68 0.179237 SYNGR4 synaptogyrin 4 -2.64 0.179237 RPA3 replication protein A3 2.07 0.179296 STXBP1 syntaxin binding protein 1 2.21 0.179296 LIPG lipase, endothelial -2.72 0.179296 KLRG1 killer cell lectin-like receptor subfamily G, member 1 2.40 0.179296 NUDCD2 NudC domain containing 2 2.08 0.179296 FAM84B family with sequence similarity 84, member B 3.16 0.179296 IRF1 interferon regulatory factor 1 -2.36 0.179296 ZNF334 zinc finger protein 334 4.81 0.179883 SLC25A1 solute carrier family 25 (mitochondrial carrier; citrate transporter), member 1 2.20 0.180060 MAN2A2 mannosidase, alpha, class 2A, member 2 -2.01 0.180101 UNC5C unc-5 netrin receptor C 2.78 0.181190 KCNT2 potassium channel, sodium activated subfamily T, member 2 -3.69 0.181454 CCBL1 cysteine conjugate-beta , cytoplasmic 2.93 0.181636 MARCH3 membrane associated ring finger 3 2.64 0.182556 TMEM40 transmembrane protein 40 -4.92 0.182630 KCNK1 potassium channel, two pore domain subfamily K, member 1 -2.23 0.183459 ITGA3 integrin alpha 3 -2.05 0.183675 RABEPK Rab9 effector protein with kelch motifs 2.75 0.183675 PTX3 pentraxin 3, long -2.63 0.183693 CLDN1 claudin 1 -3.24 0.184224 PCDHB9 protocadherin beta 9 -2.90 0.185098 ASRGL1 asparaginase like 1 2.17 0.185098 NOVA1 neuro-oncological ventral antigen 1 -3.47 0.185098 SLC1A4 solute carrier family 1 (glutamate/neutral transporter), member 4 -2.15 0.185098 PLAU plasminogen activator, urokinase -2.16 0.185098 SLC43A1 solute carrier family 43 (amino acid system L transporter), member 1 2.51 0.185361 Homo sapiens DiGeorge syndrome critical region gene 6 (DGCR6), mRNA.; protein DGCR6; Homo sapiens DiGeorge DGCR6; LOC102724770 syndrome critical region gene 6, mRNA (cDNA clone MGC:54086 IMAGE:5229172), complete cds.; Protein DGCR6 2.03 0.185639 [Source:UniProtKB/Swiss-Prot;Acc:Q14129] SLC13A4 solute carrier family 13 (sodium/sulfate symporter), member 4 -2.94 0.185696 SPANXN3 SPANX family, member N3 -3.62 0.186944 SLC35F1 solute carrier family 35, member F1 4.41 0.186944 MCTP2 multiple C2 domains, transmembrane 2 -3.56 0.187292 OSTN osteocrin 3.26 0.187429 GALNT14 polypeptide N-acetylgalactosaminyltransferase 14 4.35 0.187521 SWI5 SWI5 homologous recombination repair protein 2.84 0.187521 AFF2 AF4/FMR2 family, member 2 3.80 0.187557 FAIM2 Fas apoptotic inhibitory molecule 2 -2.35 0.187846 HLA-DRB5 major histocompatibility complex, class II, DR beta 5 -3.91 0.188078 SFXN3 sideroflexin 3 -2.12 0.188313 DDR2 discoidin domain receptor tyrosine kinase 2 2.02 0.189055 POLR3K polymerase (RNA) III (DNA directed) polypeptide K, 12.3 kDa 2.28 0.189702 ZMYM5 zinc finger, MYM-type 5 -2.12 0.189702 GABRQ gamma-aminobutyric acid (GABA) A receptor, theta -3.47 0.189935 TTC39B tetratricopeptide repeat domain 39B 2.17 0.190122 IFI44L interferon-induced protein 44-like -4.29 0.190453 COLEC12 sub-family member 12 2.19 0.190889 FLII flightless I actin binding protein -2.04 0.191209 PCDHB6 protocadherin beta 6 -4.55 0.191353 ARPC5L actin related protein 2/3 complex subunit 5-like 2.04 0.192061 LAMB3; MIR4260 laminin, beta 3; microRNA 4260 -7.53 0.192103 SMAD3 SMAD family member 3 -2.46 0.192233 TFPI tissue factor pathway inhibitor (lipoprotein-associated coagulation inhibitor) -3.55 0.192353 GSTO2 glutathione S- omega 2 -2.69 0.192431 DNAJB9 DnaJ (Hsp40) homolog, subfamily B, member 9 -2.37 0.193172 HAND1 and neural crest derivatives expressed 1 3.77 0.194813 ADAP1 ArfGAP with dual PH domains 1 2.24 0.194813 ADCYAP1R1 adenylate cyclase activating polypeptide 1 (pituitary) receptor type I 4.75 0.194813 ADGRA2 adhesion G protein-coupled receptor A2 2.06 0.194813 GALC galactosylceramidase -3.94 0.194839 BTN3A2 butyrophilin, subfamily 3, member A2 -2.05 0.195830 C16orf91 chromosome 16 open reading frame 91 2.05 0.196597 APOBEC3B apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3B -2.45 0.196597 PDIA4 protein family A, member 4 -2.03 0.196618 USP17L7 ubiquitin specific peptidase 17-like family member 7 -3.34 0.197758 SETSIP SET-like protein 2.96 0.197988 GYPC glycophorin C (Gerbich blood group) 2.96 0.197988 SCIN scinderin 2.05 0.199238 PCDHB16 protocadherin beta 16 -2.71 0.199555 CATSPER1 cation channel, sperm associated 1 -2.12 0.199555 ANKFN1 ankyrin-repeat and fibronectin type III domain containing 1 -2.35 0.200357 DLX5 distal-less homeobox 5 2.99 0.200747 Supplemental Table 2: continue

SEMA3A sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3A -2.50 0.201399 TUFT1 tuftelin 1 -2.03 0.202677 ANK1 ankyrin 1, erythrocytic -2.41 0.203372 FAM214A family with sequence similarity 214, member A -2.21 0.203372 IL32 -6.65 0.203372 SEMA4B sema domain, immunoglobulin domain (Ig), transmembrane domain (TM) and short cytoplasmic domain, (semaphorin) 4B -2.02 0.203372 UGGT2 UDP-glucose glycoprotein glucosyltransferase 2 -2.29 0.203372 CCNA1 cyclin A1 2.29 0.203372 MAP3K14 mitogen-activated protein kinase kinase kinase 14 -2.12 0.203380 BIRC3 baculoviral IAP repeat containing 3 -3.25 0.203380 AKR1B10 aldo-keto reductase family 1, member B10 () -2.06 0.203380 RASGRF2 Ras protein-specific guanine nucleotide-releasing factor 2 2.36 0.203530 KHDRBS3 KH domain containing, RNA binding, associated 3 4.20 0.203732 HIST1H3I histone cluster 1, H3i 2.83 0.204472 CDR1 cerebellar degeneration related protein 1 -2.02 0.204472 RUNX1 runt-related transcription factor 1 -2.34 0.204472 MR1 major histocompatibility complex, class I-related -2.51 0.204472 MYRF regulatory factor -2.10 0.205442 SLC35D2 solute carrier family 35 (UDP-GlcNAc/UDP-glucose transporter), member D2 -6.37 0.205971 IL7R receptor -3.62 0.206261 MARC1 mitochondrial amidoxime reducing component 1 2.15 0.206410 LRRC49 leucine rich repeat containing 49 -2.18 0.206552 TCFL5 transcription factor-like 5 (basic helix-loop-helix) 2.05 0.206700 TRIM10 tripartite motif containing 10 -2.06 0.207171 TGFB1 transforming growth factor beta 1 -2.57 0.207319 FURIN furin (paired basic amino acid cleaving enzyme) -2.33 0.208054 LIPH lipase, member H -2.11 0.208054 ANKRD18A; FAM95C ankyrin repeat domain 18A; family with sequence similarity 95, member C 2.07 0.208054 PAQR4 progestin and adipoQ receptor family member IV 2.23 0.208054 DPYD dihydropyrimidine dehydrogenase -3.99 0.209838 STS steroid sulfatase (microsomal), isozyme S 2.91 0.211094 HIST1H1T histone cluster 1, H1t -3.12 0.211155 CARD6 caspase recruitment domain family, member 6 -2.23 0.211783 LUM lumican -2.78 0.212696 SYT1 synaptotagmin I -2.54 0.212696 LBH limb bud and heart development -2.60 0.212696 DMRTA1 DMRT-like family A1 -2.69 0.214015 SERINC2 serine incorporator 2 -2.12 0.214149 ZCCHC10 zinc finger, CCHC domain containing 10 2.10 0.214175 PRKAR1B protein kinase, cAMP-dependent, regulatory, type I, beta 2.32 0.214304 VGLL3 vestigial-like family member 3 -2.92 0.216748 CYP1B1 cytochrome P450, family 1, subfamily B, polypeptide 1 -2.08 0.217849 LRP2 LDL receptor related protein 2 3.22 0.218946 TSHZ2 teashirt zinc finger homeobox 2 3.35 0.220753 EMILIN2 elastin microfibril interfacer 2 -2.58 0.220772 IL6R receptor -2.85 0.220898 BICC1 BicC family RNA binding protein 1 -2.21 0.221128 RTN4RL1 reticulon 4 receptor-like 1 3.65 0.221128 LIG4 ligase IV, DNA, ATP-dependent -2.66 0.221500 CDH23 cadherin-related 23 2.50 0.222620 GPM6A glycoprotein M6A -2.63 0.223775 ADGRE5 adhesion G protein-coupled receptor E5 -2.35 0.223775 LMX1B LIM homeobox transcription factor 1, beta 2.45 0.224040 TMEM130 transmembrane protein 130 -2.50 0.224040 TNF tumor necrosis factor -5.37 0.224040 KIAA1549L KIAA1549-like 2.39 0.225116 EPHA4 EPH receptor A4 2.27 0.225245 CCSER1 coiled-coil serine rich protein 1 2.31 0.225443 ZNF503 zinc finger protein 503 2.23 0.225601 STMN3 -like 3 2.76 0.226887 LRRC8C leucine rich repeat containing 8 family, member C -2.04 0.227782 PGR -4.12 0.227782 MMP13 matrix metallopeptidase 13 -8.58 0.227782 ZNF32 zinc finger protein 32 2.02 0.227782 GABBR2 gamma-aminobutyric acid (GABA) B receptor, 2 2.43 0.227782 TRIM55 tripartite motif containing 55 -2.06 0.227782 ANKRD18B ankyrin repeat domain 18B 2.03 0.227782 FRK fyn-related Src family tyrosine kinase -2.25 0.227782 ZDHHC22 zinc finger, DHHC-type containing 22 2.22 0.227782 RASA4 RAS p21 protein activator 4 -2.13 0.227974 COLCA2 colorectal cancer associated 2 2.06 0.228844 ARRDC3 arrestin domain containing 3 2.13 0.228864 PKIG protein kinase (cAMP-dependent, catalytic) inhibitor gamma -2.03 0.228864 ZFP36 ZFP36 ring finger protein -2.04 0.229162 KMT5C lysine (K)-specific methyltransferase 5C 2.04 0.229550 VWA5A von Willebrand factor A domain containing 5A -2.67 0.229568 SCAI; GOLGA1 suppressor of cancer cell invasion; golgin A1 2.28 0.229568 XPNPEP3 X-prolyl aminopeptidase 3, mitochondrial 2.54 0.229568 SYT14 synaptotagmin XIV -3.78 0.230916 FAM216A family with sequence similarity 216, member A 2.37 0.231097 CEACAM1 carcinoembryonic antigen-related cell adhesion molecule 1 (biliary glycoprotein) -2.04 0.231097 AR 2.21 0.231097 PCBP3 poly(rC) binding protein 3 -2.21 0.232701 SPRY2 sprouty RTK signaling antagonist 2 2.38 0.232874 RUNX2 runt-related transcription factor 2 -2.50 0.233182 MIR99AHG mir-99a-let-7c cluster host gene -2.26 0.233279 GJB7 gap junction protein beta 7 2.45 0.233571 FPGS folylpolyglutamate synthase 2.10 0.233936 SERF1B; SERF1A small EDRK-rich factor 1B (centromeric); small EDRK-rich factor 1A (telomeric) 2.03 0.234194 SERF1A small EDRK-rich factor 1A (telomeric) 2.03 0.234194 IL31RA receptor A -5.13 0.235161 Supplemental Table 2: continue

ALDH1L2 aldehyde dehydrogenase 1 family, member L2 2.36 0.235237 TLR1 toll-like receptor 1 -2.28 0.235469 ARMCX2 armadillo repeat containing, X-linked 2 -2.51 0.235892 C8orf4 chromosome 8 open reading frame 4 -2.25 0.235892 KRTAP21-2 keratin associated protein 21-2 -3.44 0.235919 ZBTB20; MIR568 zinc finger and BTB domain containing 20; microRNA 568 -3.82 0.236947 RFPL4A ret finger protein-like 4A -2.75 0.237185 SDR16C5 short chain dehydrogenase/reductase family 16C, member 5 -2.19 0.237315 FAM20C family with sequence similarity 20, member C 2.05 0.240017 DDIT3 DNA-damage-inducible transcript 3 -2.56 0.240083 PLIN2 perilipin 2 2.42 0.240226 ARL15 ADP-ribosylation factor like GTPase 15 2.17 0.240227 SYNPO synaptopodin -2.29 0.240227 ALPK3 alpha kinase 3 -2.23 0.240474 SOX11 SRY box 11 -2.41 0.241008 COL9A3 collagen, type IX, alpha 3 2.64 0.241459 SAMD9 sterile alpha motif domain containing 9 -3.80 0.241735 F2RL2 coagulation factor II (thrombin) receptor-like 2 -3.64 0.241735 ARRDC4 arrestin domain containing 4 -2.12 0.241735 CAMK1G calcium/calmodulin-dependent protein kinase IG -2.65 0.241735 LHPP phospholysine phosphohistidine inorganic pyrophosphate phosphatase -2.15 0.241735 DES -2.05 0.241735 FOSL1 FOS-like antigen 1 -2.79 0.241735 RARB , beta 3.94 0.242651 REXO1L10P REX1, RNA exonuclease 1 homolog-like 10, pseudogene -3.05 0.243730 LMNA A/C -2.01 0.244792 SLITRK5 SLIT and NTRK-like family, member 5 -2.26 0.245036 THBS1 thrombospondin 1 -2.74 0.245486 DGKA diacylglycerol kinase alpha -3.09 0.246302 FZD4 frizzled class receptor 4 2.18 0.246357 ANKRD19P ankyrin repeat domain 19, pseudogene 2.32 0.246769 SLC45A3 solute carrier family 45, member 3 -2.06 0.246971 PRR5-ARHGAP8 PRR5-ARHGAP8 readthrough 2.32 0.247324 IFIH1 interferon induced, with helicase C domain 1 -2.47 0.247324 CYP3A5 cytochrome P450, family 3, subfamily A, polypeptide 5 -2.38 0.248155 LHX2 LIM homeobox 2 2.40 0.248155 HLA-DOB major histocompatibility complex, class II, DO beta -2.27 0.248159 CCL20 chemokine (C-C motif) ligand 20 -2.52 0.252810 WFDC2 WAP four-disulfide core domain 2 -2.25 0.252810 USP17L1 ubiquitin specific peptidase 17-like family member 1 -2.46 0.253684 BEGAIN brain-enriched guanylate kinase-associated -2.46 0.254223 DBF4 Transcript Identified by AceView, Entrez Gene ID(s) 10926 2.09 0.254235 MYOM3 myomesin 3 -2.33 0.254235 PCDH10 protocadherin 10 -2.18 0.254758 C1S complement component 1, s subcomponent -2.27 0.257500 HOXA5 homeobox A5 2.62 0.258495 CSF1 colony stimulating factor 1 (macrophage) -2.09 0.258596 CXCL8 chemokine (C-X-C motif) ligand 8 -3.45 0.258596 ENDOG endonuclease G 3.89 0.262148 PLA2G7 phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma) 2.83 0.263764 RFPL4AL1 ret finger protein-like 4A-like 1 -2.74 0.264409 RENBP renin binding protein -2.80 0.265119 Supplemental Table 3: Expression levels of the different G protein subunits (α, β and γ) in the parental and ∆Gs cells. The statistical analysis (unpaired one-way ANOVA between-samples; p < 0.05 being considered statistically significant) was done using the “transcriptome analysis console” from Affymetrix. Using a threshold of a minimum of 2 fold change (<-2 or >2), none of the G-protein encoding genes was found to be differentially expressed. The false discovery rate (FDR) adjustment has been calculated for each gene and is indicated.

parental Bi-weight ∆Gs Bi-weight Fold Change ANOVA FDR Gene Description Avg Signal Avg Signal (linear) p-value p-value Symbol (log2) (log2) (parental vs ∆Gs) (parental vs ∆Gs) (parental vs ∆Gs) GNA11 guanine nucleotide binding protein (G protein), alpha 11 12.79 12.85 -1.04 0.135977 0.394487 GNA12 guanine nucleotide binding protein (G protein) alpha 12 11.64 11.22 1.34 0.126440 0.383426 GNA13 guanine nucleotide binding protein (G protein), alpha 13 11.84 12.22 -1.30 0.023307 0.211094 GNA14 guanine nucleotide binding protein (G protein), alpha 14 5.35 5.38 -1.03 0.490967 0.723730 GNA15 guanine nucleotide binding protein (G protein), alpha 15 7.75 7.64 1.08 0.630218 0.815803 GNAI1 guanine nucleotide binding protein (G protein), alpha i1 12.77 13.38 -1.52 0.034347 0.236947 GNAI2 guanine nucleotide binding protein (G protein), alpha i2 12.92 13.29 -1.30 0.000435 0.112203 GNAI3 guanine nucleotide binding protein (G protein), alpha i3 13.29 12.97 1.25 0.106507 0.355931 GNAO1 guanine nucleotide binding protein (G protein), alpha o 8.66 7.80 1.82 0.179868 0.447122 GNAQ guanine nucleotide binding protein (G protein), alpha q 10.67 9.73 1.92 0.014903 0.187846 GNAT1 guanine nucleotide binding protein (G protein), alpha t-rod 6.45 6.49 -1.03 0.823536 0.920852 GNAT2 guanine nucleotide binding protein (G protein), alpha t-cone 7.20 7.21 -1.01 0.920552 0.966923 GNAT3 guanine nucleotide binding protein (G protein), alpha t-gus 4.29 4.73 -1.35 0.312938 0.583998 GNAZ guanine nucleotide binding protein (G protein), alpha z 10.50 9.83 1.59 0.001963 0.129017 GNB1 guanine nucleotide binding protein (G protein), beta 1 15.07 15.24 -1.12 0.128356 0.386013 GNB2 guanine nucleotide binding protein (G protein), beta 2 12.73 13.33 -1.52 0.011808 0.179296 GNB3 guanine nucleotide binding protein (G protein), beta 3 7.91 7.67 1.19 0.198847 0.469394 GNB4 guanine nucleotide binding protein (G protein), beta 4 11.51 11.39 1.09 0.370855 0.631538 GNB5 guanine nucleotide binding protein (G protein), beta 5 7.53 7.98 -1.37 0.039106 0.244321 GNGT1 guanine nucleotide binding protein (G protein), gamma 1 4.98 5.45 -1.39 0.240187 0.512642 GNG2 guanine nucleotide binding protein (G protein), gamma 2 7.33 7.48 -1.11 0.511698 0.739180 GNG3 guanine nucleotide binding protein (G protein), gamma 3 4.16 4.28 -1.09 0.750803 0.884181 GNG4 guanine nucleotide binding protein (G protein), gamma 4 11.38 12.32 -1.91 0.002438 0.133236 GNG5 guanine nucleotide binding protein (G protein), gamma 5 13.84 13.82 1.02 0.503797 0.733463 GNG7 guanine nucleotide binding protein (G protein), gamma 7 9.57 9.31 1.20 0.074743 0.308821 GNG8 guanine nucleotide binding protein (G protein), gamma 8 5.77 5.83 -1.04 0.729230 0.872955 GNGT2 guanine nucleotide binding protein (G protein), gamma 9 6.14 6.69 -1.46 0.332215 0.600536 GNG10 guanine nucleotide binding protein (G protein), gamma 10 10.74 10.19 1.46 0.047841 0.262132 GNG11 guanine nucleotide binding protein (G protein), gamma 11 7.36 8.22 -1.81 0.006633 0.157845 GNG12 guanine nucleotide binding protein (G protein), gamma 12 11.50 12.22 -1.65 0.019686 0.202782 GNG13 guanine nucleotide binding protein (G protein), gamma 13 6.21 5.98 1.17 0.380751 0.638756 Supplemental Figure 1. Genomic sequences of GNAS- and GNAL-double mutant HEK293 cells. sgRNA-target sequences of the three mutant clones (A, CL1; B, CL2; C, CL3) were determined by a TA-cloning method. sgRNA target sequences are boxed and PAM sequences (NGG) are underlined. Arrows indicate a putative double-stranded break site. Restriction enzyme sites (Hap II (GNAS and GNAL #2), Pvu II (GNAL #3) and Hae III (GNAL #1)) are highlighted in red.

10 Supplemental Figure 2. Genotyping of Gs-mutant clones identified by a restriction enzyme- digested fragment method. sgRNA targets including one site (A) in the GNAS gene site and the three sites (sgRNA #1, #2 and #3, shown in B, C and D, respectively) in the GNAL gene were PCR- amplified and treated with the corresponding restriction (RE; Hap II (A and C), Hae III (B) or Pvu II (D)). The digested samples were subjected to a capillary electrograph analysis (MultiNA, Shimadzu) and pseudo-gel images of electropherogram were visualized by an accessory software with DNA markers of Hae III-digested phaiX174. Filled (grey) and open arrowheads indicate undigested and RE-digested, respectively, PCR fragments of the targeted sites in the parental HEK293 cells. An open arrowhead with an asterisk (CL2 in A) denotes a heteroduplex between the small and large single-stranded DNA. Note that there are four Hae III sites in a parental PCR fragment (B).

11 A An alignment of the Gs amino acids

GNAs-targeting sgRNA #2

B An alignment of the Golf amino acids

GNAL-targeting sgRNA #3

GNAL-targeting sgRNA #1

GNAL-targeting sgRNA #2

Supplemental Figure 3. An alignment of deduced amino acid sequences of the GNAS-(Gαs, A) and GNAL- (Gαolf, B) mutant alleles from the ∆Gs-HEK293 clones. Genomic sequences of the GNAS (A) and GNAL (B) gene from the three mutant clones (CL1, CL2 and CL3) were determined with a TA cloning method. A) Note that all alleles were introduced with a frame-shift mutation. Parent Gαs sequence refers to the isoform GNASL (long isoform). B) Note that except for one allele (CL3 allele #1, which has 28-amino acid insertion), all alleles were introduced with a frame-shift mutation. Parent Gαolf sequence refers to the isoform 2. Arrows indicate the GNAS-andGNAL-(#1, #2 and #3 constructs were used to generate the CL1, CL2 and CL3, respectively) targeting sgRNA site. Aligned regions with 100% identity are shaded in black and those above 50% identity are shaded grey.

12 epnefloigete aorsi AP 0n)o osoi Fk 10 (Fsk, Forskolin or 100nM) ∆ (AVP, vasopressin either following response ∆ epnefloigete spoeeo IO 10 (ISO, isoproterenol either following response the of of Characterization clones – 4 Figure Supplemental A esrdfloigete S rinpoeA38 1 M ciain h ubra h o fteISO the of top the Ca at number the The of activation. mM) % (10 the A23187 represent ionophore or columns ISO either following measured ...o needn xeiet,ec efre ntilct.Clm rpswr nlzdb two- by analyzed were graphs Column comparison triplicate. multiple in Bonferroni’s a performed by each followed ANOVA experiments, way independent 3 of S.E.M. C B sHK9Tclswr rninl rnfce with transfected transiently were cells Gs-HEK293T with transfected transiently cells Gs-HEK293T cAMP response (BRET2) cAMP response (BRET2) Ca2+ response (AUC x 105) 0 2 4 6 8 sHK9Tclstasetytasetdwt h PCBE isno eetse o cAMP for tested were biosensor EPAC-BRET the with transfected transiently cells ∆Gs-HEK293T HEK293 HEK293 HEK293 Overexpressed Overexpressed V2R L L CL3 CL2 CL1 Endogenous ∆Gs-HEK293 L L CL3 CL2 CL1 L L CL3 CL2 CL1 ∆Gs-HEK293 ∆Gs-HEK293 β 2AR 2+ β 2 epneotie ihA28.Dt r xrse stema ± mean the as expressed are Data A32187. with obtained response AR  )o osoi Fk 10 (Fsk, Forskolin or M) A23187 ISO 2 n PCBE isno eetse o cAMP for tested were biosensor EPAC-BRET and V2R 13 sHK9 cells. ∆Gs-HEK293 β A n blnboesr n Ca and biosensor, obelin and 2AR post-hoc test. A aetlHK9Tadthree and HEK293T Parental (A)   )atvto.()HK9Tor HEK293T (B) activation. M) )atvto.()HK9Tor HEK293T (C) activation. M) +2 epnewas response Supplemental Figure 5 – Scatter plot of the microarray comparing between parental and ∆Gs-HEK293 cells. Gene Level Differential Expression Analysis of the parental vs ∆Gs (clone 1) cells was performed on 21,448 genes using the Clarion S Human array type (Affymetrix). Among these, 491 genes were found to be either transcriptionally up-regulated (198, green) or down-regulated (293, red), including GNAL and GNAS that were knockdown using the CRISPR/Cas9 system. The list of 489 up- or down-regulated genes (with a minimum threshold of 2 fold) are presented in Supplemental Table 2. A Purinergic (P2X) Microarray signal (a.u.)

B Adenosine (P1Y) Purinergic (P2Y) Microarray signal (a.u.)

Supplemental Figure 6 – mRNA expression of P2X and P2Y purinergic receptor subtypes in HA-β2AR- HEK293S cells. Total mRNA expression in HA-β2AR-HEK293S cells was analyzed by gene expression array. Data were normalized using a quantile normalization procedure (see Methods). For comparison purposes, the expression levels of the housekeeping genes β-actin and α- are shown, since these genes are highly expressed in HA-β2AR-HEK293S cells. Since HEK293 cells have previously been found to not express Gαo to detectable amounts by Western blotting (Law et al., 1993), the normalized value for this probe is included on the graph to represent a baseline level of fluorescence of the microarray probes. The expression of the endogenous β2AR (not the overexpressed HA-β2AR, due to the location of the β2AR probe in the 3’- untralnslated region) is shown for comparison, as well as the expression of all of the purinergic receptor subtypes in HA-β2AR cells.

15