Fig. 5.

Loss of Snf5 leads to increased apoptosis, polyploidy, and poor cell growth. (a) MEFs from two wild-type embryos (blue) and three Snf5^flox/- embryos (red) were transduced with pBabe-puro^r (dotted lines, control) or with pBabe Cre-puro^r (solid lines). Two days after transduction, cells were selected in puromycin, and at day 4, cells were plated for growth assay and monitored daily by absorbance of the Wst-1 reagent. (b) Cell cycle analysis by means of BrdUrd incorporation and propidium iodide staining reveals cell cycle arrest in Snf5^flox/- Cre cells as evidenced by decreased S phase and increased G₂/M; apoptosis evidenced by increased sub-G₁ population and increased polyploidy seen as >4N population. (c) Western blot analysis demonstrating decreased levels of both Snf5 and p16Ink4a and increased levels of both p53 and p21 after deletion of Snf5.