FIGURE 2.

The depletion of MOV10 affects miRNA-mediated repression of protein translation in 293T cells. A, sequences of mir25 and mir16 and their target sites used for reporter gene constructs are shown. B, effects of MOV10-specific siRNA on the endogenous MOV10 expression in 293T cells. Values represent portions of MOV10 normalized against actin relative to control values. C and D, 293T cells were first transfected with MOV10-specific siRNA or siRNA for gfp as a control. After 24 h, cells were transfected with a plasmid containing the Renilla luciferase reporter gene with (C) perfect or (D) imperfect binding sites for mir25 or mir16 in the 3′-UTR (the plasmid also contained the firefly luciferase gene as an intraplasmid transfection normalization reporter). psiCHECK-2 was also transfected as a control. At 24 h post-transfection, Renilla/firefly luciferase activities were measured. E, 293T cells were first transfected with MOV10-specific siRNA (gfp-specific siRNA as a control). At 24 h post-transfection, cells were co-transfected with psiCHECK-2 and Renilla luciferase-specific siRNA (gfp-specific siRNA as a control). After 24 h, Renilla/firefly luciferase activity was measured. Data in C, D, and E represent mean ± S.D. (error bars). *, statistically significant, ≤0.05.