Figure 5

O -GlcNAcylation of Thr-366 and Thr-367 plays a role in PARP1 recruitment. Protein extracts from 293T cells transiently transfected with plasmids expressing K-RTA or O-GlcNAc mutants for 24 h were subjected to co-immunoprecipitation (Co-IP) assays by using M2-FLAG resins. Equal aliquots of each Co-IP were resolved on SDS-PAGE followed by western blot analysis using indicated antibodies. After normalized to respective total IP proteins (α-FLAG), the relative ratios of O-GlcNAcylation state (α-RL2) and the amount of PARP1 recruited by K-RTA and O-GlcNAc mutants are denoted beneath each lane.