Fig. 3.
Intact CRL7SMU1 complex is required for mitotic progression. (A) HeLa cells transfected with either control siRNA or SMU1-specific siRNAs were stained with propidium iodide and cell cycle analysis (whether the cells were 2N, 4N or >4N) was performed by flow cytometry. (B) HeLa cells were transfected with indicated siRNAs. The transition of cells through mitosis was analyzed by live cell time-lapse microscopy after synchronizing cells by using double thymidine block. (C) Time taken by each cell from mitotic entry to division was calculated and the data were plotted for control and SMU1-depleted cells (n=15). (D) HeLa cells were transduced with control shRNA, Cul7 shRNA, DDB1 shRNA, RNF40 shRNA and SMU1 shRNA separately. 72 h post transduction, cells were probed with antibody against phosphorylated H3 (pH3) to measure the mitotic index. Nuclei were counterstained with DAPI. (E) Quantification of mitotic index results. Error bars indicate the mean+s.d.; *P<0.05, **P<0.01