Fig 2: BAP1 depletion leads to mitotic progression defects and loss of SAC integrity.A MSTO-211H and NCI-H2452 were either mock-depleted or depleted of BAP1 or BRCA1 as described in Fig. 1A. Cells were fixed and stained with Hoechst 33258 to observe the DNA and histograms show the percentage of mitotic cells in the different mitotic phases indicated. B Cells treated and stained as in A, were analysed to measure the percentage of metaphase cells with misaligned chromosomes (means ± S.D.) based on n = 3 independent experiments (60 mitotic cells for each condition). C, D The percentage of interphase cells treated as in A that were multinucleated (C) or contained micronuclei (D) were scored. E Cell lysates treated as in A were analysed by Western blot with antibodies against of MAD2L1 and a-tubulin. F Cells treated as in A were processed for immunofluorescence microscopy with BUBR1 antibodies and DNA stained with Hoechst 33258. Histograms show the mean intensity of BUBR1 on metaphase chromosomes following BAP1 or BRCA1 depletion relative to that observed upon mock depletion. Data in A, B, C, D and F are expressed as means ± S.D. (n = 3).