Fig. 6

NR6A1 modulates miR-205-5p expression in HepG2 cells. (a) HepG 2 cells were transfected with scrambled or NR6A1 siRNA for 48 h. The expressions of a set of miRNA genes potential targeting INSR were checked by qPCR method. n = 3. **P < 0.01. (b) Gel electrophoresis of PCR product of miR-205-5p in control and NR6A1 silenced HepG2 cells. Representative image from 3 independent experiments was shown. (c & d) miRNA-205-5p mimic was transfected in HepG2 cells for 48 h. Oil Red O staining was performed to examine intracellular lipid content. Representative images were shown. Bar = 100 μm. n = 3. *P < 0.05. (e & f) DGAT2, FAS, PEPCKc, CPT1a, MCAD, SCD and INSR (f) mRNA levels in HepG2 cells in the presence or absence of 50 nM miR-205-5p mimics. n = 3. **P < 0.01., *P < 0.05. (g) Immunoblot analysis was performed to detect expressions of phosphorylated INSR, total INSR and phosphorylated p70-S6K in HepG2 cells which cotransfected with NR6A1 siRNA and miR-205-5p mimic. Representative images were shown. n = 3. (H) HepG2 cells were transfected with luciferase reporter plasmids containing the INSR 3’UTR combined with miR-205-5p mimic or miR-NC. n = 3. HepG2 cells were transfected with either scrambled or NR6A1 siRNA alone, or cotransfected with INSR or AKT siRNA for 48 h. (i) miR-205-5p expression was detected by qPCR method. (j) Representative gel electrophoresis of PCR products was shown. n = 3. **P < 0.01., *P < 0.05., ns. no significant