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Fig. 3. NER response to local Hoechst 33342 treatment + 405 nm irradiation. (A) XPC-GFP-expressing cells were irradiated through a filter with UV-C light (spots indicated by arrows), sensitized with Hoechst 33342 and subsequently locally treated with 405 irradiation in the nucleus (lines indicated by arrowheads). Induction of CPDs is shown by the CPD antibody staining (right panel) both on UV-C and H+405 treated areas. XPC-GFP accumulated on both areas irradiated through a filter with UV-C light (arrows) and irradiated with 405 nm in combination with Hoechst 33342 (arrowheads). (B) Treatment as in panel A, here cells were stained with an antibody that recognizes 6-4PPs (right panel). Surprisingly, no 6-4PP-staining can be detected on laser-irradiated areas (lines indicated by arrowheads), while the UV-C treated areas show a clear induction (arrows). The bar graph indicates fluorescence intensities of the nucleus (1), 405 nm combined with Hoechst 33342 treatment induced local damage (2) and UV-C induced local damage (3). (C) GFP-XPA accumulates to a low level on local damage induced by 405 nm laser irradiation in combination with Hoechst 33342 treatment (arrowhead) compared with local UV-C irradiated areas (arrow). The bar graph indicates fluorescence intensities of the nucleus (1), 405 nm combined with Hoechst 33342 treatment induced local damage (2) and UV-C induced local damage (3).
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