Fig. 2. SpII*, FANCA and XPF co-localize to discrete nuclear foci after treatment of normal cells with 8-MOP plus UVA light. Normal human lymphoblastoid cells were either undamaged or treated with 8-MOP plus UVA light (6 kJ m^-2) and the localization of SpII*, FANCA and XPF in the nucleus examined 15 hours after treatment with 8-MOP plus UVA. (A) Dual staining was carried out using affinity-purified monoclonal anti--spectrin antibody and affinity-purified polyclonal anti-FANCA antibody, and stained cells were analyzed by immunofluorescence. When the fluorescent signals for SpII* (green) and FANCA (red) are merged, the overlapping foci are yellow, indicating co-localization of these two proteins. (B) An analysis similar to that in (A) was carried out using anti--spectrin antibody (green) and affinity-purified polyclonal anti-XPF antibody (red). Fluorescent signals for both proteins were merged and overlapping foci appear yellow. (C) Dual staining was also carried out using anti-FANCA (green) and anti-XPF (red) antibodies. The fluorescent signals were merged and overlapping foci are yellow. In all of the above experiments, cells were also stained with the appropriate preimmune sera.