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. 2014 Aug 13;9(8):e104821. doi: 10.1371/journal.pone.0104821

Figure 4. Effects of RITA in PCI-13 (p53 null) cells expressing mutant p53.

Figure 4

(A) PCI-13 (p53 null) cells expressing vector control or the indicated p53 constructs were treated with RITA 2.5 µM for 72 hours after which protein lysates were prepared and levels of total and phosphorylated p53 and p21 were assessed by western blotting. β-actin was used as an internal loading control. (B) PCI-13 cells expressing the indicated constructs were seeded on 6-well plates, treated with RITA at the indicated concentrations, and counted in a clonogenic assay. With the exception of pBabe cells treated at 0.25 µM, all cell lines at all tested concentrations of RITA exhibited significantly decreased colony formation compared to untreated control (p<0.05). (C) PCI-13 cells expressing the indicated constructs were seeded in 6-well tissue culture plates and treated with 0.25 µM of RITA after which cells were fixed and stained for senescence-associated -β-galactosidase (SA-β-gal) activity. Representative images from three experiments with similar results are shown. All cell lines exhibited significantly increased SA-β-gal staining compared to untreated control (p<0.05).