SUPPLEMENTARY FIG. S2. b-galactosidase staining of
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1 SUPPLEMENTARY FIG. S1. b-galactosidase staining of senescent cells in 500 mg=dl glucose (10 magnification). SUPPLEMENTARY FIG. S3. Toluidine blue staining of chondrogenic-differentiated adipose-tissue-derived mesenchymal stem cells lineage in 100 mg=dl (10 magnification). Note the difference in the pellet size as well as the cell=matrix density. SUPPLEMENTARY FIG. S2. b-galactosidase staining of senescent cells in 500 mg=dl glucose þ tumor necrosis factoralpha treatment (10 magnification). SUPPLEMENTARY FIG. S4. Toluidine blue staining of chondrogenic-differentiated adipose-tissue-derived mesenchymal stem cells lineage in 500 mg=dl of glucose (10 magnification). Note the difference in the pellet size as well as the cell=matrix density.
2 SUPPLEMENTARY FIG. S7. Oil Red O staining of adipogenic-differentiated adipose-tissue-derived mesenchymal stem cells in 250 mg=dl glucose concentrations (10 magnification). SUPPLEMENTARY FIG. S5. Representative experiment of 4 genes expressed in chondrogenic-differentiated nascs and dascs. All values were normalized to 100 nascs (x-axis; n ¼ 3). dasc, diabetic adipose-tissue-derived mesenchymal stem cell; nasc, nondiabetic adipose-tissue-derived mesenchymal stem cell. SUPPLEMENTARY FIG. S6. Oil Red O staining of adipogenicdifferentiated adipose-tissue-derived mesenchymal stem cells in 100 mg=dl glucose (10 magnification).
3 SUPPLEMENTARY FIG. S8. Regression analysis comparing nasc and dasc (100 mg=dl) gene expression. The data were normalized based on nascs. dasc, diabetic adipose-tissue-derived mesenchymal stem cell; nasc, nondiabetic adiposetissue-derived mesenchymal stem cell. Red and green circles indicate up regulation and down regulation of genes, respectively. Black circles indicate of genes displaying less than two fold difference between analyzed groups.
4 SUPPLEMENTARY FIG. S9. Regression analysis of nasc (250 mg=dl) genes as a function of glucose. The data were normalized based on nascs 100 mg=dl. nasc, nondiabetic adipose-tissue-derived mesenchymal stem cell. Red and green circles indicate up regulation and down regulation of genes, respectively. Black circles indicate of genes displaying less than two fold difference between analyzed groups.
5 SUPPLEMENTARY FIG. S10. Regression analysis of nasc (250 mg=dl) genes as a function of insulin. The data were normalized based on nascs 250 mg=dl without insulin. nasc, nondiabetic adipose-tissue-derived mesenchymal stem cell. Red and green circles indicate up regulation and down regulation of genes, respectively. Black circles indicate of genes displaying less than two fold difference between analyzed groups.
6 SUPPLEMENTARY FIG. S11. Regression analysis of dasc (250 mg=dl) genes as a function of glucose. The data were normalized based on dascs 100 mg=dl. dasc, diabetic adipose-tissue-derived mesenchymal stem cell. Red and green circles indicate up regulation and down regulation of genes, respectively. Black circles indicate of genes displaying less than two fold difference between analyzed groups.
7 SUPPLEMENTARY FIG. S12. Regression analysis of dasc (250 mg=dl) genes as a function of insulin. The data were normalized based on dascs without insulin. dasc, diabetic adipose-tissue-derived mesenchymal stem cell. Red and green circles indicate up regulation and down regulation of genes, respectively. Black circles indicate of genes displaying less than two fold difference between analyzed groups.
8 Supplementary Table S1. List of Analyzed Genes Gene Full name GenBank ABCC8 ATP-binding cassette, sub-family C (CFTR=MRP), member 8 NM_ ACE angiotensin I converting enzyme (peptidyl-dipeptidase A) 1 NM_ ACLY ATP citrate lyase NM_ ADRB3 adrenergic, beta-3-, receptor NM_ AGT angiotensinogen (serpin peptidase inhibitor, clade A, member 8) NM_ AKT2 v-akt murine thymoma viral oncogene homolog 2 NM_ AQP2 aquaporin 2 (collecting duct) NM_ CCL5 chemokine (C-C motif ) ligand 5 NM_ CCR2 chemokine (C-C motif ) receptor 2 NM_ CD28 CD28 molecule NM_ CEACAM1 carcinoembryonic antigen-related cell adhesion molecule 1 (biliary NM_ glycoprotein) CEBPA CCAAT=enhancer binding protein (C=EBP), alpha NM_ CTLA4 cytotoxic T-lymphocyte-associated protein 4 NM_ DUSP4 dual specificity phosphatase 4 NM_ ENPP1 ectonucleotide pyrophosphatase=phosphodiesterase 1 NM_ FBP1 fructose-1,6-bisphosphatase 1 NM_ FOXC2 forkhead box C2 (MFH-1, mesenchyme forkhead 1) NM_ FOXG1 forkhead box G1 NM_ FOXP3 forkhead box P3 NM_ G6PC glucose-6-phosphatase, catalytic subunit NM_ G6PD glucose-6-phosphate dehydrogenase NM_ GCG glucagon NM_ GCGR glucagon receptor NM_ GCK glucokinase (hexokinase 4) NM_ GLP1R glucagon-like peptide 1 receptor NM_ GPD1 glycerol-3-phosphate dehydrogenase 1 (soluble) NM_ GSK3B glycogen synthase kinase 3 beta NM_ HMOX1 heme oxygenase (decycling) 1 NM_ HNF4A hepatocyte nuclear factor 4, alpha NM_ ICAM1 intercellular adhesion molecule 1 NM_ IDE insulin-degrading enzyme NM_ IFNG interferon, gamma NM_ IGFBP5 insulin-like growth factor binding protein 5 NM_ IKBKB inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta NM_ IL10 interleukin 10 NM_ IL12B interleukin 12B (natural killer cell stimulatory factor 2, cytotoxic lymphocyte NM_ maturation factor 2, p40) IL4R interleukin 4 receptor NM_ IL6 interleukin 6 (interferon, beta 2) NM_ INPPL1 inositol polyphosphate phosphatase-like 1 NM_ INSR insulin receptor NM_ IRS1 insulin receptor substrate 1 NM_ PDX1 pancreatic and duodenal homeobox 1 NM_ INS insulin NM_ IRS2 insulin receptor substrate 2 NM_ MAPK14 mitogen-activated protein kinase 14 NM_ MAPK8 mitogen-activated protein kinase 8 NM_ ME1 malic enzyme 1, NADP(þ)-dependent, cytosolic NM_ NEUROD1 neurogenic differentiation 1 NM_ NFKB1 nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 NM_ NOS3 nitric oxide synthase 3 (endothelial cell) NM_ NRF1 nuclear respiratory factor 1 NM_ NSF N-ethylmaleimide-sensitive factor NM_ PARP1 poly (ADP-ribose) polymerase 1 NM_ PIK3C2B phosphoinositide-3-kinase, class 2, beta polypeptide NM_ PIK3CD phosphoinositide-3-kinase, catalytic, delta polypeptide NM_ PIK3R1 phosphoinositide-3-kinase, regulatory subunit 1 (alpha) NM_ PPARA peroxisome proliferator-activated receptor alpha NM_ PPARG peroxisome proliferator-activated receptor gamma NM_ PPARGC1A peroxisome proliferator-activated receptor gamma, coactivator 1 alpha NM_ PPARGC1B peroxisome proliferator-activated receptor gamma, coactivator 1 beta NM_ PRKAA1 protein kinase, AMP-activated, alpha 1 catalytic subunit NM_ PRKAG2 protein kinase, AMP-activated, gamma 2 non-catalytic subunit NM_ (continued)
9 Supplementary Table S1. (Continued) Gene Full name GenBank PRKCB1 protein kinase C, beta NM_ PTPN1 protein tyrosine phosphatase, non-receptor type 1 NM_ PYGL phosphorylase, glycogen, liver NM_ RAB4A RAB4A, member RAS oncogene family NM_ RETN resistin NM_ SELL selectin L NM_ SLC2A4 solute carrier family 2 (facilitated glucose transporter), member 4 NM_ SNAP23 synaptosomal-associated protein, 23kDa NM_ SNAP25 synaptosomal-associated protein, 25 kda NM_ SREBF1 sterol regulatory element binding transcription factor 1 NM_ STX4 syntaxin 4 NM_ STXBP1 syntaxin binding protein 1 NM_ STXBP2 syntaxin binding protein 2 NM_ HNF1B HNF1 homeobox B NM_ TGFB1 transforming growth factor, beta 1 NM_ NKX2-1 NK2 homeobox 1 NM_ TNF tumor necrosis factor (TNF superfamily, member 2) NM_ TNFRSF1A tumor necrosis factor receptor superfamily, member 1A NM_ TRIB3 tribbles homolog 3 (Drosophila) NM_ VAMP3 vesicle-associated membrane protein 3 (cellubrevin) NM_ VAPA VAMP (vesicle-associated membrane protein)-associated protein A, 33 kda NM_ VEGFA vascular endothelial growth factor A NM_003376
10 Supplementary Table S2. Transcriptomic Alterations of nascs and dascs in Response to Increased Glucose Concentration and Insulin Treatment Genes dascs vs. nascs (100 mg=dl) dascs vs. nascs (250 mg=dl) nascs (100 vs. 250 mg=dl) dascs (100 vs. 250 mg=dl) nascs vs. nascs þ Ins in 250 mg=dl dascs vs. dascs þ Ins in 250 mg=dl CTLA INSR ND 1.60 ND CEACAM ND SELL 2.78 ND 2.47 ND ADRB3 ND SLC2A4 ND ND SNAP25 ND ND STXBP2 ND ND ND ND GCK ND ND GPD ND FBP ND IFNG IL ND ND ND IL ND IL12B ND RETN ND VEGFA ND ND ND DUSP ND 2.76 CEBPA ND ND FOXP ND ND ND HNF4A ND ND PDX ND 1.93 ND HNF1B ND PPARGC1A ND ND NKX2-1 ND ND 4.96 List of genes obtained from comparison between nascs and dascs grown in glucose condition of 100 and 250 mg=dl with and without insulin treatment (n ¼ 6). Positive values represent an up-regulation in the dascs. Ratios are greater than 2(P < 0.05). ND, not detected
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