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. 2004 Sep 23;23(19):3874–3885. doi: 10.1038/sj.emboj.7600375

Figure 6.

Figure 6

Phosphorylation of Xrcc4 Thr233 is sensitive to CK2 inhibition in vivo. (A) XR-1 cells stably expressing Xrcc4250Δ and Xrcc4250ΔT233A were incubated in the presence of 75 μM TBB or DMSO for 4 h. WCEs were then examined by anti-phosphothreonine (upper panels) or anti-Xrcc4 immunoblotting (lower panels). (B) WCEs from the Xrcc4250Δ XR-1 cell line treated as in (A) were mixed with PNKFHA in a PD assay, followed by immunoblotting with anti-V5 antibody (to detect V5-tagged Xrcc4250Δ) and anti-GST antibody. (C) HEK293T cells transiently expressing V5-tagged PNK were treated with 75 μM TBB for 4 or 24 h, or DMSO as a control, harvested and the WCEs were immunoprecipitated with anti-V5 antibody. The WCEs and immunoprecipitates were then examined by anti-Xrcc4, anti-V5 (PNK) immunoblotting as indicated.