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. 2015 Apr 21;35(10):1741–1753. doi: 10.1128/MCB.01475-14

FIG 2.

FIG 2

PATZ1 is downregulated upon DNA damage. (A) Downregulation of PATZ1 in DOX-treated MEFs. Whole-cell lysates from MEFs treated with 1 μM DOX for the indicated times were immunoblotted with anti-PATZ1 (top) and anti-p53 (bottom) antibodies. The presence of PATZ1 isoforms and p53 is indicated by arrows. (B) Downregulation of PATZ1 alternative splice variants in HCT116 (p53 WT or p53−/−) cells treated with 1 μM DOX for the indicated times. (Top and middle) Whole-cell lysates were immunoblotted with anti-PATZ1 (top) and anti-p53 (middle) antibodies. (Bottom) Equal loading was confirmed by blotting of the same membrane with antiactin antibodies, revealing equal amounts of a 40-kDa actin band. (C) DOX treatment does not alter mRNA levels of PATZ1. PATZ1 mRNAs from HCT116 p53 WT or HCT116 p53−/− cells treated with 1 μM DOX for the indicated times were quantified by real-time PCR. PATZ1 expression was normalized to Gapdh expression. (D) Loss of PATZ1 activates p53 function. Quantitative RT-PCR analysis reveals higher p53-dependent gene transcript levels at time points where PATZ1 is lost upon DNA damage. Mock-infected cells express more p21 (left) and Puma (right) than do PATZ1-overexpressing cells after 48 h of DOX treatment.