miR-33b targets c-Myc in osteosarcoma cells.

(A) The sequences of miR-33b binding sites within the human c-Myc 3′UTRs and schematic reporter constructs, in this panel, c-Myc-WT represent the reporter constructs containing the entire 3′UTR sequences of c-Myc. C-Myc-MUT represent the reporter constructs containing mutated nucleotides. (B) The analysis of the relative luciferase activities of c-Myc-WT, c-Myc-MUT in 293T cells. The error bars are derived from triplicate expriments. (C) qRT-PCR analysis of c-Myc mRNA expression in MG-63 cells after treatment with miRNA mimics or scramble or untreated. The expression of c-Myc was normalized to GAPDH. (D)Western blot analysis of c-Myc expression in MG-63 cells transfected with miR-33b mimics or scramble or untreated. GAPDH was also detected as a loading control. (E) The ratio signal of c-Myc to GAPDH in each lane was determined. Student’s t test was used to analyze the significant differences. ***p<0.001.