Figure 2: FTO promotes adipogenesis via mitotic clonal expansion.

(a) Representative image showing BrdU incorporation, 24 h after induction of adipogenic differentiation in MEFs from WT and FTO-KO mice, BrdU (green), DAPI (blue); scale bar, 100 μM. (b) Quantification analysis of BrdU incorporation in WT versus FTO-KO MEFs. (c) Representative image showing BrdU incorporation, 24 h after induction of adipogenic differentiation in MEFs from WT and FTO-4 mice, BrdU (green), DAPI (blue); scale bar, 50 μM. (d) Quantification analysis of BrdU incorporation in WT versus FTO-4 MEFs. (c,d) Data represent six experiments using MEFs derived from one mouse of each genotype. (e,f) qPCR of mRNA for the indicated genes in FTO-4 MEFs treated with control (black) or FTO (green) siRNA before (e) or 48 h after (f) adipogenic induction. Data are expressed relative to expression of GAPDH. Data represent three experiments each with three technical replicates on MEFs from one mouse. (g,h) qPCR of mRNA for CCND1 (g) and CCND3 (H) in FTO-4 MEFs treated with control (black) or FTO (green) siRNA, at 16, 24 and 40 h after induction of adipogenic differentiation. Data represent 1 (0 h) or 4 (16, 24 and 40 h) biological replicates each with three technical replicates. Independent Students’ t-test (c,d), multivariant ANOVA (e,f), repeated-measures ANOVA (g,h). *P<0.05; ***P<0.001 against WT (c,d) or control (e–h). Data presented in graphs represent means±s.e.m.