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. 2013 Jan 25;20(5):721–731. doi: 10.1038/cdd.2012.169

Figure 5.

Figure 5

USP7 depletion causes accumulation of multipolar mitosis and accumulation of Aurora-A kinase. (a) Immunofluoresce staining of HEp2 and H1299 cell lines expressing shRNAs using Aurora-A and α-tubulin to label spindle poles. Hoechst was used to stain DNA. The panels on the right represent the quantification of multipolar events over the total number of mitosis (±S.D., n=3). Analysis was conducted counting at least 100 mitotic events for each experiment. (b) Immunofluorescence staining of Aurora-A, α-tubulin and DNA in HEp2 and H1299 stably expressing USP7 shRNA. In USP7-depleted cells, we observed multipolar divisions with overnumerary poles (n>4). Quantification of these events is presented on the right panels (±S.D., n=3). (c) Representative western blot analysis showing accumulation of Aurora-A in HEp2 and H1299 cells depleted by control or USP7. Cells were synchronized by DTB, and released in growth media for 11 h to progress through mitosis. Numbers represent quantification of Aurora-A protein over internal control (actin)