Fig. 5.

Inhibition of Kpnβ1 results in increased p53 stability and reporter activity, as well as increased p21 and decreased Mcl-1 in response to cisplatin. a, b SiHa cells were treated with DMSO or 5 μM INI-43 for 2 h (a) or transfected with si-ctrl or si-Kpnβ1 for 48 h (b) followed by 50 μg/mL CHX treatment. Protein was harvested at the indicated time points, and p53 content analyzed by western blot. GAPDH served as the loading control. The full-length blots are shown in Supplementary Fig. 5A and 5B. c The fold increase in p53 half-life is shown as the mean ± SEM from the three independent experiments (*p < 0.05). d p53 reporter activity is increased upon 24 h 5 μM INI-43 treatment of SiHa cells (*p < 0.05). Experiments were performed in triplicate and repeated at least three independent times. e p53 reporter activity is enhanced in INI-43-cisplatin combination treated SiHa cells, compared to cells treated with cisplatin alone (*p < 0.05). Experiments were performed in triplicate and repeated at least three independent times. f Western blot showing levels of p53 after single and combination treatment. β-tubulin served as a loading control. The full-length blots are shown in Supplementary Fig. 5C. g Western blot showing levels of p53 targets p21 and Mcl-1 after single and combination treatment. β-tubulin served as a loading control for Mcl-1 and GAPDH for p21. Results shown are representative of two independent experiments. The full-length blots are shown in Supplementary Fig. 5D