Fig. 4.
Reduced accumulation of XPAH244R impairs ERCC1-XPF recruitment at UV lesions. (A) Representative images of the recruitment of indicated GFP-tagged XPA variants to UV-C laser tracks, measured by live cell imaging. (B) Quantification of (A). Data represents average and SD of three independent experiments of 25 to 78 cells per experiment. (C) Representative images of the recruitment of indicated GFP-tagged XPA mutants reconstituted in U2OS(FRT) XPA-KO cells, to sites of damage after 30 J/m2 UV-C irradiation through 5-µm pore membranes. Sites of local damage are identified by TFIIH/p89 staining. (D) Quantification of (C). Quantification of the recruitment of TFIIH/p89 is shown in SI Appendix, Fig. S5B. (E) Representative images and (F) quantification of ERCC1 recruitment in the indicated primary fibroblasts after 30 J/m2 UV-C irradiation through 5-µm pore membranes. Sites of local damage are identified by TFIIH/p89 staining. Quantification of the recruitment of TFIIH/p89 is shown in SI Appendix, Fig. S5C. (G) Representative images and (H) quantification of ERCC1 recruitment in the indicated U2OS(FRT) cells after 30 J/m2 UV-C irradiation through 5-µm pore membranes. Sites of local damage are identified by TFIIH/p89 staining. Quantification of the recruitment of TFIIH/p89 is shown in SI Appendix, Fig. S5D. (D, F, and H) All cells are depicted as individual data points with the bar representing the mean of all data points. The individual means of three biological replicates are depicted as colored points with black circles.