Figure 10

Inhibition of PARG results in enhanced recruitment EGFP-UL31 to sites of laser microirradiation. (A) Cells expressing EGFP-UL31 were treated with 0.1% DMSO (vehicle) or 1 μM DEA, a PARG inhibitor, two hours prior to imaging. Images of cells were captured immediately before (Pre), immediately after (Post) and 30 seconds after the region within the white circle was microirradiated using a 405 nm laser line. The intensity of radiation and size of the area of irradiation were maintained between all cells irradiated. (B) We define the ratio of areas, Area^Recruit/Area^Rad, as the area of protein recruitment at Δmax (Area^Recruit) divided by the initial area that was microirradiated (Area^Rad). (C) Graph of the ratio of areas of UL31, NBS1 and H2A1.1 macro domain in DMSO (vehicle) and 1 µM DEA treated cells. Area^Recruit/Area^Rad was calculated for each construct (n ≥ 11) from two independent experiments. Error bars represent standard error of the mean. (ns = not significant, *p < 0.05, ***p < 0.001).