Fig 2: DDX24 mediated inhibition of cancer cell growth is independent of p53.((a) Left panel) BJ-hTERT, HCT116, MDA-MB-231, and PC-3 cell lines were transfected with 2.5 nM pooled DDX24 siRNAs (GE Healthcare) in quadruplicates. Cell number, representation, and statistical tests as in Fig. 3 (P-values: 0.0026, 0.0004, and 0.0002, respectively). ((a) Right panel) BJ-hTERT, HCT116, and PC3 cells were transfected with 5 nM of pooled DDX24 siRNA (GE Helthcare) and 4 individual siRNAs (Qiagen) in 6 replicates. Cell number, representation, and statistical tests as in Fig. 3 (P-values < 0.0001). (b) HCT-116 with p53 wild-type (p53+/+) and p53 null (p53−/−) were transfected with 5 nM of control or DDX24 siRNAs and plated for colony formation assay. Cells were transfected in triplicate and plated at 1,000 cells per well followed by quantification of total colony area. Bars represent average +/− 1 SD (P-values: 0.0005 and <0.0001, respectively). (c) Western blot analysis of p53, p21, and DDX24 expression in wild-type and p53−/− HCT-116 cells. Cells were transfected with 5 nM of siRNAs and lysed 72 hrs post-transfection followed by WB with corresponding antibodies. (d) Western blot analysis of p21 and DDX24 expression in DU145 and BJ-hTERT cells. Cells were transfected with 5 nM of pooled siRNAs and lysed 72 hrs post-transfection followed by WB with corresponding antibodies.