Figure 6. miR-96-5p regulated p66shc expression in hepG2 cells. HepG2 cells were transfected with ago-96-5p to upregulate miR-96-5p expression. Ago-NC was used as a normal control. (A, B) Argonaute-2 (Ago2) – immunoprecipitated miR-96-5p expression in NAFLD mice and PA-treated hepG2 cells. (C) miR-96-5p expression. (D, E) P66shc and cyto C protein expressions. (F) P66shc mRNA expression. (G) Schematic of the wild-type p66shc 3′UTR (3′UTR-wt) and mutated p66shc 3′UTR (3′UTR-mut) luciferase constructs. (H) HepG2 cells were transfected with 3′UTR-wt or 3UTR-mut and with ago-96-5p or ago-NC, as indicated. **P< 0.05. Error bars depict the standard deviation. BS: binding site; NC: negative control.