Molecules and Cells

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Fig. 4.

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Fig. 4. (A–C) HeLa cells were transfected with a siControl or siCbl for 72 h. The cells were cotreated with a separase siRNA (siSeperase) for 72 h or 100 nM BI 2536, as a Plk1 inhibitor, for 2 h. Images are maximum projections from Z-stacks of representative cells stained for Centrin-1 (green), β-tubulin (red), and DNA (blue). The level of proteins were analyzed by western blotting using the indicated antibodies (B). Metaphase cells with multipolar spindles or pseudobipolar spindles were quantified and plotted (C; n = 100 cells for each quantification). (D) Seventy-two hours after siRNA transfection, HeLa cells were fixed in MeOH and stained for Centrin-1 (green), C-NAP1 (red), and DNA (blue). G2 cells with multiple or disengaged centrioles were quantified and plotted (n = 100 cells for each quantification). (E) HeLa cells were transfected with siRNAs and prepared as in . Metaphase cells with multipolar spindles were quantified and plotted (n = 100 cells for each quantification). (F and G) HeLa cells were transfected with a siControl or siCbl. The cells were fixed in MeOH and stained with antibodies as indicated. The Sas6, STIL, and PLK4 intensities were quantified and plotted (n = 100 centrioles for each quantification). Alternatively, the cells were harvested at 72 h post-transfection and lysates were analyzed by western blotting using the indicated antibodies (G). (H) Seventy-two hours after siRNA transfection, the mRNA levels of c-Cbl and DDA3 in nocodazole-treated mitotic cells were quantified by RT-qPCR analysis (n = 3). AU, arbitrary units. Data are represented as mean ± SEM. Scale bars = 5 μm. *P < 0.01.
Mol. Cells 2019;42:840~849 https://doi.org/10.14348/molcells.2019.0142
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