Figure 2

Influence of ENOblock on the adipogenic program in differentiating preadipocytes and mitochondrial membrane potential. (A) Schematic of the compound treatment protocol in primary cultures of differentiating white adipocytes. (B) Effect of 72 h treatment with 10 µM forsoklin, 1 µM rapamycin,10 µM ENOblock or 1 mM NaF on the expression of adipogenesis regulatory genes in differentiating adipocytes. (C) Expression of oxidative phosphorylation regulatory genes. (D) Expression of thermogenesis regulatory genes. (E) Live cell imaging of TMRE fluorescence to visualize mitochondrial membrane potential in 3T3-L1 white preadipocytes and brown preadipocytes after treatment with 10 µM ENOblock, 1 mM NaF, 10 µM forsoklin or 1 µM rapamycin for 72 h. (F) Quantification of mitochondrial membrane potential in 3T3-L1 preadipocytes. (G) Quantification of mitochondrial membrane potential in brown preadipocytes. (H) Oil red O staining of 3T3-L1 white preadipocytes treated with 10 µM ENOblock, 1 mM NaF, 10 µM forsoklin or 1 µM rapamycin for 72 h and adipogenic factors for 5 days. (I) Quantification of oil red O staining in the treated adipocytes. n = 9; ns: not significantly different. *, ** or ***: significantly different from the corresponding ‘Control’ or ‘Untreated’ respectively with p < 0.05, p < 0.01 or p < 0.001; ## or ###: significantly different from the corresponding ‘NaF’ sample with p < 0.01 or p < 0.001; ж, жж or жжж: significantly different from the corresponding ‘ENOblock’ sample respectively with p < 0.05, p < 0.01 or p < 0.001.