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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
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    UNIVERSITA’ DEGLI STUDI DI PARMA Dottorato di ricerca in Fisiopatologia Sistemica Ciclo XX ROLE OF GLYPICAN-6 AND NG2 AS METASTASIS PROMOTING FACTORS Coordinatore: Chiar.mo Prof. Ezio Musso Tutor: Chiar.mo Prof.Roberto Perris Dottoranda: Katia Lacrima Anni Accademici 2005-2008 To Indy L'anima libera e' rara, ma quando la vedi la riconosci: soprattutto perché provi un senso di benessere, quando gli sei vicino. (Charles Bukowski ) Index Summary ……………………………………………………..................................................... 3 1. Introduction …………………………………………………………………………………… 5 1.1. Proteoglycans (PGs)………………………………………………………………......... 6 1.2. Membrane associated proteoglycans……………………………………………..…... 8 1.3. Syndecans……………………………………………………………………………..…. 9 1.4. Glypicans……………………………………………………………………………...….. 11 1.5. GPC6……………………………………………………………………………….…….. 13 1.6. NG2/CSPG4……………………………………………………………………….…….. 14 1.7. Metastasis………………………………………………………………………….….…. 16 1.8. Soft Tissue Sarcoma (STS)…………………………………………………………….. 17 1.9. Membrane PGs and tumour……………………………………………………………. 18 1.10. Membrane PGs in sarcoma…………………………………………………………….. 24 2. Material and Methods ……………………………………………………………………… 26 2.1. Cell Culture……………………………………………………………………….………. 27 2.2. RNA extraction……………………………………………………………………….…. 28 2.3. Real Time quantitative PCR……………………………………………….………….. 28 2.4. DNA extraction…………………………………………….……………………………. 30 2.5. Plasmids and Transfection………………………………………….………………… 30 2.6. Western Blotting………………………………………………………………………... 31 2.7. Preparation of ECM substrates………………………………………….……………
  • Investigation of Candidate Genes and Mechanisms Underlying Obesity

    Investigation of Candidate Genes and Mechanisms Underlying Obesity

    Prashanth et al. BMC Endocrine Disorders (2021) 21:80 https://doi.org/10.1186/s12902-021-00718-5 RESEARCH ARTICLE Open Access Investigation of candidate genes and mechanisms underlying obesity associated type 2 diabetes mellitus using bioinformatics analysis and screening of small drug molecules G. Prashanth1 , Basavaraj Vastrad2 , Anandkumar Tengli3 , Chanabasayya Vastrad4* and Iranna Kotturshetti5 Abstract Background: Obesity associated type 2 diabetes mellitus is a metabolic disorder ; however, the etiology of obesity associated type 2 diabetes mellitus remains largely unknown. There is an urgent need to further broaden the understanding of the molecular mechanism associated in obesity associated type 2 diabetes mellitus. Methods: To screen the differentially expressed genes (DEGs) that might play essential roles in obesity associated type 2 diabetes mellitus, the publicly available expression profiling by high throughput sequencing data (GSE143319) was downloaded and screened for DEGs. Then, Gene Ontology (GO) and REACTOME pathway enrichment analysis were performed. The protein - protein interaction network, miRNA - target genes regulatory network and TF-target gene regulatory network were constructed and analyzed for identification of hub and target genes. The hub genes were validated by receiver operating characteristic (ROC) curve analysis and RT- PCR analysis. Finally, a molecular docking study was performed on over expressed proteins to predict the target small drug molecules. Results: A total of 820 DEGs were identified between
  • Interleukin 2 Medical Intensive Care Unit (4MICU)

    Interleukin 2 Medical Intensive Care Unit (4MICU)

    Interleukin 2 Medical Intensive Care Unit (4MICU) Ronald Reagan UCLA Medical Center 757 Westwood Plaza Los Angeles, CA 90095 Main Phone: (310) 267-7441 Fax: (310) 267-3785 About Our Unit The Medical Intensive Care Unit (MICU) cares Quick for critically ill patients in an intensive care Reference Guide environment, with nursing staff specially trained in the administration of Interleukin 2 therapy. Unit Director / Manager Mark Flitcraft, RN, MSN One registered nurse (RN) is assigned to take (310) 267-9529 care of a maximum of two patients. Our Medical Clinical Nurse Specialist Intensive Care Unit patient rooms are designed Yuhan Kao, RN, MSN, CNS (310) 267-7465 to allow nurses constant visual contact with their patients. As a safety precaution, the Medical Assistant Manager Sherry Xu, RN, BA, CCRN Intensive Care Unit is a closed unit and requires (310) 267-7485 permission to enter by intercom. Clinical Case Manager Each private-patient-care room contains the Connie Lefevre (310) 267-9740 most advanced intensive-care equipment available, including cardiac-monitoring and Clinical Social Worker Codie Lieto emergency-response equipment. The curtains in (310) 267-9741 the room will usually be drawn to keep your room Charge Nurse On-Duty more private. (310) 267-7480 or (310) 267-7482 A brief tour is available on weekdays for patients and visitors interested in walking through the unit Patient Affairs (310) 267-9113 and meeting the staff before arrival. To arrange for a tour, please call the nurse manager at Respiratory Supervisor (310) 267-9529. Orna Molayeme, MA, RCP, RRT, NPS (310) 267-8921 UCLAHEALTH.ORG 1-800-UCLA-MD1 (1-800-825-2631) About Our Unit During Your Stay Quick The Medical Team Reference Guide During each shift, you will be assigned a registered nurse (RN) and a clinical care partner (CCP).