Co-localization of WDR76 with DNA Repair Proteins to Damaged Sites.

<p>(A) HEK293FRT cells stably expressing Halo-WDR76 and transiently expressing CLIP-PARP1, XRCC5 and RPA1 were stained with Hoechst dye, micro-irradiated with a 405 nm UV laser. Merged images of ligand labeled proteins after laser microirradiation are also provided. In cases where there is more than one cell in image the two cells were not damaged simultaneously. (B) Kinetics of recruitment to microirradiated regions of WDR76 (pink lines and symbols), XRCC5 (red), PARP1 (black) and RPA1 (blue). Cells were imaged every second, and intensity values were binned over 5-s intervals. Microirradiation was initiated at time = 0s. Averages are shown with the standard error. (C) Kinetics of recruitment to microirradiated regions of PARP1, XRCC5, RPA1 and WDR76 represented as box plots with the box showing the standard error with a coefficient of 1 and the whiskers showing the standard deviation with a coefficient of 1. (D) Enrichment of Halo-WDR76 at sites of localized DNA damage induced by UV laser microirradiation. HEK293FRT cells stably expressing Halo-WDR76 were stained with Hoechst dye, micro-irradiated with a 405 nm UV laser, fixed, and analyzed by indirect immunofluorescence using anti-γ-H2AX. The yellow line indicates site of co-enrichment.