Figure 3. HDAC inhibitors and DNA damage agents regulate GLS2 expression through TAp63 induction. (A) H1299 cells were treated with TSA (1 μM) or LBH589 (2 μM) for 18 h, and whole-cell extracts were analyzed by IB using antibodies to the indicated proteins (left panels). In parallel total RNA was extracted, and GLS2 expression was analyzed by qRTR-PCR (right panels). (B) H1299 cells were transfected twice with siRNA oligos targeting a non-relevant mRNA or p63 mRNA. After 48 h of each siRNA oligo transfection, cells were treated with HDAC inhibitors as described in (A). Cells were used either for quantification of GLS2 mRNA by qRT-PCR (right panel) or subjected to IB analysis utilizing antibodies to the indicated proteins (left panel). H1299 (C) and Hep3B (D) cells were treated with 1 μM doxorubicin for the indicated time points. Total RNA was extracted and used to measure GLS2 and TAp63 mRNA levels by qRT-PCR. Data shown are the mean ± SD of three replicates.