FIG 3.

mir-21 partially mediates RASSF1A regulation of SKP2 and cell cycle progression. (A to C) HeLa and U2OS cells were cotransfected with the indicated oligonucleotides. Whole-cell lysates were immunoblotted for detection of the indicated proteins. (B and C) HeLa cells were treated as described for panel A. (D) HeLa cells were transfected with the indicated oligonucleotides and treated with nocodazole (100 ng/ml) for 16 h prior to staining with propidium iodide and FACS analysis. Bars indicate cell populations gated for 2N (G₁), >2N and <4N (S), and 4N (G₂/M) DNA content. (E) HeLa (left) and HBEC30 (right) cells were transfected as indicated followed by incubation with BrdU for 5 h (HeLa) or 20 h (HBEC30). Bars indicate mean relative BrdU incorporation ± SEM. ***, P < 0.001; **, P < 0.01, Student's t test. (F) HeLa cells were transfected with indicated siRNAs, and whole-cell lysates were immunoblotted for detection of the indicated proteins.