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. 2014 Jan 23;9(1):e86358. doi: 10.1371/journal.pone.0086358

Figure 2. Epistatic analysis for Ku80 and Pol β.

Figure 2

All cells are deleted for p53 (even controls) to avoid early replicative senescence. Shown is the average of three experiments. (A) Western showing RNAi knockdown of Pol β in p53-/- control and ku80-/- p53-/- fibroblasts that stably express a shRNA plasmid specific for mouse Pol β (three clones). PCNA was loaded to normalize for nuclear protein levels. The expression of endogenous Pol β in the shRNA-transduced cell lines was undetectable by immunoblot, consistent with our earlier reports for this mouse Pol β -specific shRNA [30]. (B) Dose-response to MMS for p53-/- control and ku80-/- p53-/- fibroblasts with and without mouse Pol β shRNA expression. (C) Western showing increased Pol β levels for the ku80-/- p53-/- fibroblasts that stably express a Pol β expression plasmid (two clones). Beta-actin was loaded to normalize for cellular protein levels. (D) Pol β -overexpression rescues Ku80-mutant phenotype for MMS.