Fig. 4
From: PARP2 mediates branched poly ADP-ribosylation in response to DNA damage
The recruitment of PARP2 to DNA lesions is mediated by PARP1-dependent PARylation. a The recruitment kinetics of PARP2 to DNA lesions. GFP-PARP1 or PARP2 were expressed in U2OS cells. Following laser micro-irradiation treatment, the recruitment of PARP1 or PARP2 was examined with live-cell imaging at the indicated time points (left panel). The relocation kinetics is shown in the right panel. Data represent mean ± s.d. from three biologically independent experiments (right panel). At least 20 cells were included in each experiment. b The recruitment of endogenous PARP2 was examined following laser micro-irradiation by anti-PARP2 antibodies. c The NTR domain alone is sufficient to be recruited to the DNA damage sites. The recruitment kinetics of GFP-NTR at different time points was measured. d PARP1 is required for the recruitment of PARP2. GFP-PARP2 or the NTR domain was expressed in the Parp1+/+ or Parp1−/− MEFs. The recruitment kinetics was examined at indicated time points. The percentage of recruitment under different PARP1 conditions is shown in the right panel e. The catalytic activity of PARP1 is required for the recruitment of PARP2. PARP1-deficient U2OS cells were stably expressing the full length PARP1 or the E988A mutant (catalytic inactive mutant). GFP-PARP2 was expressed in the cells, and the recruitment kinetics was measured. Scale bar represents 5 μm
