Fig. 4.

USP48 is recruited to ICLs and attenuates clearance of γH2AX foci. a Workflow for the recruitment of proteins to ICLs in U2OS and VU1131 cells. b Localization of FANCD2 and GFP-USP48 (WT and C98S mutant) to sites of laser micro-irradiation after Trioxalen treatment in U2OS and VU1131 cells. Scale bar = 10 μm. c Representative immunofluorescence images after staining for γH2AX on WT, ΔUSP48, ΔFANCC, and ΔUSP48ΔFANCC cells after treatment with MMC (30 nM) for 18 h and then recovery for the indicated time-points. Scale bar = 10 μm. d Quantification of γH2AX foci of the indicated cell lines. UT untreated cells. Quantification was performed using the Cell Profiler software. Error bars show mean with S.D. (standard deviation). Statistical significance was determined by Wilcoxon test with p < 0.05 as threshold. N.S. not significant; **** = p < 0.0001