Fig. 5

NR6A1 regulates INSR expression in HepG2 cells. (a) The mRNA expression of INSR in NR6A1 silenced HepG2 cells. n = 3. **P < 0.01. (b) HepG2 cells were transfected with scrambled or NR6A1 siRNA fragments for 24 h, and then starved in low serum (0.5% FBS) containing medium overnight before stimulated with insulin for another 15 min. Immunoblot analysis was performed to detect expressions of phosphorylated INSR and total INSR. Representative blots were shown. n = 3. (c) HepG 2 cells were infected with Ad-LacZ or Ad-NR6A1 for 48 h. The mRNA expression of INSR was analyzed by using qPCR method. n = 3. *P < 0.05. (d) Immunoblot analysis was performed to detect expressions of phosphorylated INSR and total INSR in Ad-NR6A1 infected cells. Representative blots were shown. n = 3. HepG2 cells were transfected with either scrambled or NR6A1 siRNA alone, or cotransfected with INSR siRNA for 48 h. (e) Total protein was collected for western blotting assay. Representative blots from 3 independent experiments were shown. (f) Oil Red O staining was performed to show the lipid accumulation. Representative images were shown. Bar = 100 μm. (g) Oil Red O contents in HepG2 cells. n = 3. **P < 0.01., *P < 0.05