Fig. 3

Regulation of lipid accumulation by NR6A1 is mTORC1 dependent. (a) HepG2 cells were transfected with control or NR6A1 siRNA fragments for 48 h, SREBP1c and SREBP2 transcripts were analyzed by using qPCR assay. n = 3. (b) HepG2 cells were transfected with scrambled, PEPCKc, SREBP1c or Rictor siRNA fragments for 48 h. The gene silencing was confirmed by using qPCR assay. n = 3. **P < 0.01. (c) Representative images of Oil Red O staining in HepG2 cells which transfected with NR6A1 siRNA alone or co-transfected with PEPCKc, SREBP1c or Rictor siRNA. Bar = 100 μm. n = 3. (d) Quantification by extracting Oil Red O stained lipid droplets with 100% isopropanol. n = 3. *P < 0.05. The phosphorylation of S6K in NR6A1 silenced- (e) or overexpressed (f) HepG2 cells were analyzed by using western blotting. Representative blots were shown. n = 3. (g) The control or NR6A1 silenced cells were treated with rapamycin for 16 h. Oil Red O staining was performed to show lipid content. Representative images were shown. Bar = 100 μm. n = 3. (h) Oil Red O contents in HepG2 cells. n = 3. *P < 0.05. ns. no significant