Figure 2

Acute loss of Rb leads to chromosome instability and increases MPM2 positive cells. (A) pRb deficient MEFs show increased aneuploid metaphases both hypodiploid (<40 chromosomes) and hyperdiploid (>40 chromosomes). Each bar indicates mean + s.e. of three independent experiments. (B) pRb deficient MEFs have increased percentage of cells with micronuclei. An average of 100 cells were analyzed for each time point, each bar represents mean + s.e. of two experiments. (C) MEFs acutely depleted of pRb (Rb-/-) have increased number of mitotic cells with multiple centrosomes (arrowheads). Mitotic cells and centrosomes were revealed by using primary antibodies against MPM2 (Mitotic Protein Monoclonal) and γ-tubulin, that were detected by FITC (green) and TRITC (red) conjugated secondary antibody respectively. (D) Histograms indicating the percentage of MPM2 positive cells in pRb competent and deficient MEFs. An average of 100 cells were analyzed for each time point, each bar represents mean + s.e. of at least two experiments. (E) Western blot analysis showing the amount of MAD2 protein after 3 days from the adenoviral infection: Rb^LoxP/LoxP MEF uninfected (lane 1), infected with the empty adenoviral vector (lane 2) and with the Ad-Cre vector (lane 3).