Fig. 4

The effect of NR6A1 on insulin signaling in HepG2 cells. The control or NR6A1 silenced cells (a) and Ad-LacZ or Ad-NR6A1 infected cells (b) were cultured overnight in low serum (0.5% FBS) containing medium followed by treatment with insulin (10 nM or 100 nM) for 15 min. Cellular lysates were immunoblotted for AKT, phospho-AKT, mTOR, phospho-mTOR and phospho-GSK3β. Representative blots were shown. n = 3. (c) HepG2 cells were transfected with either scrambled or Rictor siRNA for 48 h. The mRNA expression of Rictor was analyzed by qPCR method. n = 3. (d) NR6A1 siRNA was cotransfected with either control siRNA or Rictor siRNA for 24 h, and starved overnight in low serum (0.5% FBS) medium before treated with insulin (10 nM) for 15 min. The expression of AKT and phospho-AKT were checked by western blotting. Representative blots were shown. n = 3