FIG. 5.

Mir-17-5p regulates translation of endogenous E2F1. Cell lysates (50 μg) from MCF-7 (A) and ZR-75-1 (D) were separated by SDS-PAGE, transferred onto nitrocellulose membrane, and probed with anti-AIB1 and anti-E2F1 antibody. The same blot was stripped and reprobed with anti-β-actin antibody and used as a loading control (bottom panel). (B and C) MCF-7 cells were transfected with Mir-17-5p, Mir-95, or vehicle only. Cells were starved in methionine-free medium and then pulse-labeled with [³⁵S]methionine. The cells were then harvested and immunoprecipitated with anti-E2F1 antibody. Precipitated proteins were separated by SDS-PAGE and visualized by using a phosphorimager.