Killing Cells with Chromosomal Instability Figure 4. P53-dependent and p53-independent apoptosis. (a–d) Dotted line shows the en.CD8GFP marked test region and the other half expressed no transgenes. Acridine Orange staining on double (a–b: Candidate and mad2) and triple knockdown (c–d: Candidate, mad2 and p53) wing discs. (e) Graph shows quantitation of Acridine Orange staining (above wild type) in control and candidate knocked down imaginal wing disc halves. The first bar of each group represents candidate RNAi alone (control), the second bar represents candidate RNAi with p53RNAi (P53), the third or black bar represents Candidate and mad2 knocked down and the fourth bar represents triple knockdown (Candidate, mad2 and p53). Error bars represent 95%CIs, n$8 in all cases. P-values are calculated by two-tailed t-tests with Welch’s correction: p,0.001 = www and p.0.05 = ns (not significant). Tests compare candidate mad2 p53 with candidate alone to test whether significant p53-independent cell death is seen when each candidate is co- depleted with Mad2. Significant levels of p53-independent cell death are seen for asp, mad2 but not Pask, mad2. doi:10.1371/journal.pone.0047447.g004 identify mechanisms by which cancer cells can tolerate the adverse effects of CIN and aneuploidy which in turn may lead to the identification of novel targets that can specifically kill advanced, drug resistant, CIN tumor cells without harming normal cells. PLOS ONE | www.plosone.org 7 October 2012 | Volume 7 | Issue 10 | e47447