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. 2021 Jun 24;9:693079. doi: 10.3389/fcell.2021.693079

FIGURE 6.

FIGURE 6

The SUMOylation reduced the activity of PPARγ2 through K107 site and weakened adipogenic differentiation. (A) The SUMOylation of PPARγ2 was relied on Lys residues of 107 and 395. Flag-PPARγ2, Flag-PPARγ2-K107R, ubc9 and RH-SUMO3 were co-expressed in HEK293T cells and co-IP assays were performed with Flag and RGS antibody. (B) The expression of PPARγ2 in ST2 cells transfected by Flag-PPARγ2 for 24h (a) and NC-siRNA and PPARγ2-siRNA for 36 h (b) were detected by Western Blot. (C) NC-SiRNA, PPARγ2–SiRNA, Flag-PPARγ2, and Flag-PPARγ2-K107R were transfected in mouse mesenchymal stromal cell line ST2 cells and the expression of adipogenic marker genes C/EBPα (a), adiponectin (b), and FABP4 (c) was analyzed by Q-PCR after 7-day adipogenic induction. (D) Flag-PPARγ2, Flag-PPARγ2-K107R, and RH-SUMO3 were transfected in ST2 cells and the expression of adipogenic marker genes C/EBPα (a), adiponectin (b), and FABP4 (c) was analyzed by Q-PCR after 7-day adipogenic induction. Data are shown as the mean ± SEM. **p < 0.01, ***p < 0.001; ****p < 0.0001. All the data were obtained from at least three independent experiments.