Fig. 4

SGK3 was a target of miR-182 and negatively regulated by miR-182 in PCOS GCs and KGN cells. (A) The potential binding sites between miR-182 and SGK3 were exhibited. (B and C) PCOS GCs and KGN cells were co-transfected with SGK3-WT or SGK3-MUT and miR-con, miR-182, in-miR-con or in-miR-182 and then the luciferase activity was measured via dual-luciferase reporter assay. (D and E) The enrichment of SGK3 and miR-182 in Anti-Ago2 or Anti-IgG immunoprecipitation complex in PCOS GCs and KGN cells was detected by RIP and qRT-PCR assays. (F) The level of miR-182 in PCOS GCs and KGN cells transfected with miR-con or miR-182 was detected by qRT-PCR. (G and H) The protein level of SGK3 in PCOS GCs and KGN cells transfected with miR-con, miR-182, in-miR-con or in-miR-182 was examined by western blot assay. *P < 0.05