Figure 7.

Adipocyte differentiation is inhibited in G0s2-knockout MEF cells, and ectopic expression of G0s2 in G0s2-knockout MEF restores adipogenesis. (a) MEFs from wild-type (WT) or G0s2-knockout (KO) mice were induced for adipocyte differentiation. Expression of G0s2 was analyzed by reverse transcriptase-PCR, and microscopy was done at day 12. (b) Western blotting analysis of G0s2 and adipocyte-specific proteins in WT and KO MEFs. (c) KO MEFs were transfected with G0s2-FLAG-overexpression vector. Western blotting analysis of expression in WT containing empty vector (WT), G0s2-KO containing empty vector (KO), or rescued G0s2-KO (KO-G0s2) was performed with anti-FLAG antibody. (d) BODIPY staining of WT, G0s2-KO, and KO-G0s2 MEF cells. The fluorescent dyes BODIPY and DAPI (4,6-diamidino-2-phenylindole) were used to stain the lipids contained in droplets of differentiated fat cells and nuclei of existing cells. (e) WT or KO MEFs were transfected with siRNA directed against ATGL, and microscopy ( × 20) was performed. (f) Western blotting analysis was carried out in WT or KO MEFs after ATGL siRNA transfection