Fig. 6

PPARγ promotes beige cell differentiation of MSCs by binding the UCP1 promoter. A After IRISIN stimulation for different durations, p-PPARγ expression in cells was detected by western blotting. B After IRISIN stimulation for different durations, the cell lysate was prepared by nucleolysis. The expression of p-PPARγ in the nucleus was detected by western blotting. C After IRISIN stimulation for 7 days, PPARγ phosphorylation was detected in the nucleus using a microscope. Compared to the control group, the level of phosphorylated PPARγ was higher in the nucleus of the IRISIN-treated group. D, E In the presence or absence of IRISIN stimulation, the two groups of cells were fixed with formaldehyde, and the fragment was sonicated; the PPARγ antibody captured the binding fragment and PCR was used to detect the binding fragment. F, G The UCP1 promoter was introduced into the PGL4.10 vector (PGL4.10-UCP1 promoter). PGL4.74 was used as the internal reference vector. PGL4.10-UCP1 promoter and PPARγ overexpression vectors were co-transfected into 293T cells. The control cells were co-transfected with PGL4.74 and PPARγ vectors. PPARγ overexpression was detected by the dual luciferase assay. The experiment was repeated three times. The results of dual luciferase were analyzed using primer5. The results revealed significant interaction between PPARγ and the UCP1 promoter (P < 0.05). *p < 0.05, **p < 0.01, and ***p < 0.001