Fig. 6.

G9a suppresses PTEN expression to promote adipogenesis and inhibit osteogenesis. DNMT3B and H3K9me2 are abundant at the promoter of upstream 123–329 bp PTEN locus in BM-MSCs compared with DP-MSCs. a ChIP analysis for determining the occupancy of PTEN promoter by DMNT3B and repressive marks. Below: Quantification of the results. Western blotting (b, c) and immunofluorescence (d) showed that BM-MSCs increased in the protein level of H3K9Me2 and master regulator, G9a. Bar = 25 μm. e Lentiviral knockdown of G9a in BM-MSCs increased PTEN and decreased AKP phosphorylation. f G9a knockdown cells were induced for adipogenic differentiation for 2 weeks and stained by nile red staining. Bar = 25 μm. Right Quantification of nile red positive cell percentages. g The cells were also induced for osteogenic differentiation for 3 weeks and stained by Arsenazo III kit. Bar = 200 μm. Right Quantification of the calcium contents relative to DNA contents. Results are expressed as the mean ± SD of three independent experiments