Figure 5.

p53 contributes to DCF-mediated cytotoxicity in TE11 cells. TE11 or KYSE150 cells were treated for 48 h with 200 or 400 µM DCF, respectively. RNA level of p53 was evaluated by qPCR relative to vehicle-treated control cells in TE11 in (a) or KYSE150 in (b). (c) In TE11 cells, RNA level of indicated p53 target genes was evaluated by qPCR relative to vehicle-treated control cells. (d) Protein level of p53 in TE11 cells was determined by immunoblotting 3 days following transfection of siRNA oligos targeting p53 or non-targeting siRNA oligos. β-Actin was used as a loading control. TE11 cells transfected with indicated siRNA oligos were treated with 200 µM DCF for 72 (e) or 48 (f, g) h. Cells were assessed for RNA level of TP53 by qPCR in (e), and apoptosis by Annexin V/PI flow cytometry with representative FACS dot plots shown in (f) with dot plots summarizing data from three independent experiments in (g). Data in (a, b, d, c, e, g) shown as mean ± SEM; *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001 by t-test; n = 3.