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. 2004 Jan 20;101(5):1247–1252. doi: 10.1073/pnas.0307765100

Fig. 3.

Fig. 3.

Effect of dominant negative PP5.GFP on phosphorylation of DNA–PKcs. (A) Cellular localization in unfixed cells of either GFP in subclone PP5.C13 (Left) or PP5.GFP fusion protein in subclone PP5.C7 (Right). (B) The PP5.C7 subclone expressing dominant negative PP5.GFP was irradiated with 11.5 Gy and recovered for 30–410 min. Nuclear extracts, Western blotting, and staining with anti-pT2609 antibody were part of the experiment series shown in Fig. 2; the HeLa wild-type cells of Fig. 2 A thus serve as control. * and ** denote phosphorylated DNA–PKcs degradation products. (C) Knock-down of PP5 by RNA interference. HeLa cells were transfected with siRNAs targeted against PP5. As control, HeLa cells were either mock treated or transfected with unspecific siRNAs. As before, nuclear extracts were prepared from undamaged cells and cells irradiated with 10 Gy after 30-min recovery time and stained with anti-pS2056 antibody and anti-PP5 antibody.