Figure 3.

Bat3 is necessary for DNA damage-induced p53 acetylation. (A) Normal p53 phosphorylation. U2OS cells were transfected with siCNT or siBat3 RNA oligo-duplexes and treated 48 h later with Dox (0.4 μg/mL). Protein samples were prepared at the indicated time points post-Dox. Phosphorylation of p53 on Ser15, Ser20, and Ser392 (P-p53) was detected using phospho-specific antibodies. (B) Impaired DNA damage-induced p53 acetylation. U2OS cells were transfected and treated as in A. Extracts were immunoprecipitated with anti-p53 antibody (DO-1) and immunoblotted with antibodies specific for acetylated Lys320, Lys373, and Lys382 (Ac-p53). (C) Rescue of the impaired DNA damage-induced p53 acetylation by the ΔRNAi-B3. U2OS cells were subjected to Bat3 KD and 24 h later they were transfected with WT-B3 and ΔRNAi-B3. The cells were then treated with Dox as in A. Protein levels of acetylated p53 were examined as in B. Data shown are representative of four independent preparations.