Figure 5.

Activation of p53 signaling and induction of apoptosis in TP53 wild type cell lines. (A) TP53 mutated TC-71 cell line and TP53 wild-type PDX-EW#4-C cells were treated with 3µM MC3343 for 24h. Whole cell lysates were analyzed using western blot. Three independent experiments were performed and one representative immunoblot of p-ATM, p21, phospho-p53, p53, PARP and caspase-3 cleaved is shown (B) Representative flow cytometry scatter plots showing apoptosis of p53 mutated (TC-71 and A-673) and p53 wild-type (LAP-35 and PDX-EW#4-C) EWS cells stained with Annexin-V/7-AAD following 48h treatment with MC3343 (3-5 µM) or DMSO. Numbers at the corners represent the percentage of cells found in each quadrant (viable-lower left, early apoptotic-lower right, late apoptotic-upper right, and dead cells-upper left are indicated).