Fig. 4. FRAP analysis of UV-C-treated cells expressing XPC-GFP. (A) FRAP analysis of untreated (green line) and UV-irradiated cells (blue lines) at different UV doses. (B) UV-dose-dependent and time-dependent immobilization of XPC-GFP. Percentage of UV-induced immobilization is plotted against time for the different UV-doses, non-damage-induced immobilization was set at zero. (C) Scheme of the FRAP-FLIP procedure on locally damaged areas. A small strip covering half of the local damage and spanning the entire nucleus is bleached at relatively low laser intensity for a period of 2 seconds. Subsequently fluorescence is monitored at regular time intervals in the bleached (FRAP) and non-bleached (FLIP) half of the local damage. (D) Confocal images of a locally irradiated cell expressing XPC-GFP (5 µm pore filter). Left panel, before bleaching; middle panel, directly after bleaching and right panel, 90 seconds after bleaching. (E) The relative fluorescence of the FRAP and FLIP area is shown over time. The log of fluorescence redistribution difference between FLIP and FRAP areas are plotted against time (dotted line). (F) Simultaneous FRAP/FLIP analysis of local damage at 37°C and 27°C. (G) Assembly kinetics of XPC-GFP to locally damaged areas at 37°C and 27°C. The curves are normalized to the bound fraction in the locally damaged area.