RABL6A knockdown causes centrosome amplification in WT and p53−/− MEFs.

<p>(A) Representative confocal images taken under identical settings of <i>p53</i>−<i>/</i>− CON, kd1 and kd2 cells stained for centrosomes (γ-tubulin, green) and nuclei (DAPI, blue). Yellow arrows point to areas with centrosomes, white arrows indicate micronuclei. Dashed line in the kd2 image demarcates two different cells. Scale bar, 20 µm. (B) Quantification of immunofluorescence data from (A) measuring the percent of cells with <u>></u> 3 centrosomes per cell. *, p<0.05 versus CON. (C) Immunoblot showing effective mouse RABL6A depletion in p53−/− MEFs, as well as high level overexpression of human RABL6A, with γ-tubulin levels as loading control. (D and E) Quantification of immunofluorescence data for centrosome amplification (D) and multinucleation (E) in CON, kd1 and kd2 p53−/− MEFs expressing empty vector (V) or human RABL6A (R) rescue viruses. The mean values plus standard deviations were obtained from more than 3 independent experiments (*, p<0.001 compared to CON; **, p<0.01 for human RABL6A expressing cells compared to vector controls). Statistical significance for data in panels B, D and E was determined using paired, two-tailed Student’s t-test.