Obesity-Induced Methylation of Osteopontin Contributes to Adipogenic Differentiation of Adipose-Derived Mesenchymal Stem Cells

Figure 3

OPN deficiency accelerates ASC differentiation into adipocytes. (a) ASCs from CD mice or from HFD mice were cultured in adipogenic differentiation medium for 4 days and stained with Oil Red O to reveal lipid droplets. (b) CD and HFD ASCs, transfected with DNMT1 siRNA (siDNMT1) or control siRNA (siNC), were cultured in adipogenic differentiation medium for 3 days and stained with Oil Red O to reveal lipid droplets. (c) Wild-type or OPN^−/− ASCs were cultured in adipogenic differentiation medium for 3 days and stained with Oil Red O to reveal lipid droplets. (d) Wild-type ASCs transfected with OPN siRNA (siOPN) or control siRNA (siNC) were analyzed for OPN mRNA and protein 3 days after transfection, to verify knockdown efficiency. (e) Two groups of ASCs were subjected to adipogenic differentiation medium for 6 days and stained with Oil Red O. Adipocyte size was quantification analyzed by an imaging system (ImageJ, National Institutes of Health). (Bar = 50 μm; adipocytes counted in four random microscopic fields from two wells per group.) Values are means ± SEM. ; ; . All experiments were repeated at least thrice.

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