Figure 3.

The C Terminus of CHD2 Is Required for DNA Damage Recruitment and PAR Binding

(A) Schematic representation of CHD2 and its deletion mutants.

(B) Western blot showing expression of the mutants from A in U2OS cells.

(C) Recruitment of CHD2-GFP deletion mutants in cells depleted for endogenous CHD2 by siCHD2-69 or siCHD2-17 in case of CHD2^952–1391-GFP. CHD2^1–1611-GFP was rendered siCHD2-69-resistant. NBS1-mCherry was a DNA damage marker.

(D) Quantification of (C). 10–30 cells were analyzed from two independent experiments.

(E) IP of the indicated CHD2-GFP fragments from HEK293 cells followed by southwestern blotting to monitor association with radiolabeled PAR. Recombinant PARP1 was a control. CoIP with endogenous PARP1 is shown in the bottom panel.

(F) Quantification of (E) and two additional independent experiments. PAR binding levels of CHD2^WT-GFP were set to 1.