Supplemental Data Article's title: Purinergic receptor transactivation 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 enzyme 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 genes 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 gene 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 ankyrin 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 transcription factor 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 protein 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 oncostatin M 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 membrane protein 3 -2.41 0.112203 SERPINF1 serpin 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 antigen -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 nuclear receptor 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 interleukin 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 tyrosine 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 growth factor 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, interferon-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 activating protein 2 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 tumor necrosis factor receptor superfamily, member 9 -16.22 0.122868 FILIP1L filamin 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 G protein-coupled receptor, class C, group 5, member C -2.09 0.125061 ZBTB7B zinc finger 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 myosin 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 serine peptidase inhibitor, Kunitz type 1 -3.30 0.125061 MYL9 myosin light chain 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 internexin neuronal intermediate filament protein, alpha 6.31 0.129017 CXCL10 chemokine (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 ligase 2.29 0.136366 HBP1 HMG-box transcription factor 1 -2.06 0.137726 POU6F2 POU class 6 homeobox 2 3.42 0.138586 ACTBL2 actin, 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 neurofilament, 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 keratin 17, type I -2.56 0.143625 SERPINB8 serpin peptidase inhibitor, clade B (ovalbumin), 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 interleukin 13 receptor, alpha 2 -2.55 0.149863 FLT1 fms-related tyrosine kinase 1 2.48 0.150351 ETV5 ets variant 5 -5.05 0.151176 PAMR1 peptidase domain containing associated with muscle regeneration 1 -3.90 0.153221 PLTP phospholipid transfer protein 7.87