Fig 5. Let-7c targets CDC25A in HCC cells.

(A) Firefly luciferase reporter vectors containing the CDC25A wild-type (pmiR-CDC25A-3′-UTR-wt) or mutant (pmiR- CDC25A-3′-UTR–mut) 3′-UTR were generated and co-transfected into HepG2 cells along with either the let-7c agomir or negative control to identify CDC25A targets. The 3′UTR of CDC25A mRNA contained two complementary sites for the seed region of let-7c. (b) Wild: wild-type; Mut: mutated. The seed sequence is underlined. (B) Relative luciferase activity was analyzed after the reporter plasmids or control reporter plasmid were co-transfected with let-7c into HEK-293 cells. Representative experiments are shown. (C) Western blot assays of the endogenous CDC25A protein level in HepG2 cells transfected with the let-7c agomir, negative control or let-7c inhibitor. (D) Real-time PCR assay of CDC25A mRNA expression in HepG2 cells transfected with the let-7c agomir or negative control.