Silencing of p53 expression results in reduced colony formation and elevated cell death

<p><b>Copyright information:</b></p><p>Taken from "Cancer-derived p53 mutants suppress p53-target gene expression—potential mechanism for gain of function of mutant p53"</p><p></p><p>Nucleic Acids Research 2007;35(6):2093-2104.</p><p>Published online 7 Mar 2007</p><p>PMCID:PMC1874625.</p><p>© 2007 The Author(s)</p> ( and ) p53 expression was silenced as described in HUH-7 and T47D cells using pSuper-based control or -specific siRNA, or were left untransfected. Cells were selected for two weeks on G418. Parallel cultures were used for semi-quantitative RT-PCR analysis of expression status (representative results shown for HUH-7 cells) (A) and colonies were visualized by staining with crystal violet solution (B, upper panel). All experiments were performed at least thrice independently and representative results are shown. Colonies were counted both manually and using colony count software. Similar results were obtained using both methods. Data are presented as the mean ± SD error of the mean. () Cell death was analysed by measuring the sub-G1 DNA content reflective of apoptotic cells, by flow cytometry. HUH-7 cells were transfected with scrambled or p53 siRNA (100 or 200 nM) and the percentage cell death was determined 5 days after transfection. Representative results are shown