Fig. 2: Knockdown and pharmacological inhibition of TIP60 reduces ΔNp63α acetylation in cisplatin-resistant cells.

A A431 Parental and A431 Pt cells and B A431 Pt and JHU006 were transfected with non-silencing control (NSC) and si-RNA against TIP60 (siTIP60) as indicated. C A431 Pt and JHU006 cells were treated with either DMSO as a control (Veh, vehicle) or TIP60 inhibitor NU9056 at 100 μM and 86 μM dose, respectively. D A431 Parental and A431 Pt and JHU006 cells were treated with either DMSO as a control (Veh) or 25, 50 or 75 μM doses of TIP60 specific inhibitor TH1834 for 24 h as indicated. E A431 Parental and A431 Pt cells were transiently treated for 6 h with either DMSO as a control (Veh) or 50 μM dose of TIP60 specific inhibitor TH1834 as indicated and were harvested immediately. In all panels, cells were pre-treated with HDAC inhibitors 1 μM of Trichostatin A and 5 mM of Nicotinamide for 6 h prior to immunoprecipitation (IP). Whole-cell lysate cells were immunoprecipitated with an anti-acetyl-lysine (Ac-K) antibody followed by immunoblot analysis (top panels) using antibodies specific for p63, TIP60 or β-actin. β-actin was included as a loading control for equivalent protein in each IP and Input lane. Representative immunoblots are shown. Densitometric analysis (top panel) showing the fold change in acetylated-ΔNp63α relative to control condition after normalization to input β-actin. Error bars indicate mean +1SEM from three or more independent experiments. *p ≤ 0.05 relative to the corresponding Parental (A), NSC (B) or vehicle control (C, D).