HES1 affects FANCA and FANCL nuclear localization. (A) Immunoprecipitation of FANCA and FANCC in HeLa cells treated with the γ-secretase inhibitor L685,458 (10 μM, 5 hours) or DMSO (control) and Western blotted for FANCA, FANCC, FANCG, and HES1. (B) Immunofluorescence staining of endogenous FANCA in Hes1^−/− and wild-type fibroblasts cells with and without treatment with L685,458 (10 μM, 5 hours) or DMSO control. (C) Immunofluorescence staining of endogenous FA proteins, as indicated, or HES1 in HeLa cells treated with the γ-secretase inhibitor L685,458 (10 μM, 5 hours) or DMSO (control). The staining was visualized by confocal microscopy. Confocal fluorescence settings were established with untreated cells. (D) FA protein levels from HeLa cell extracts treated with L685,458 (10 μM, 5 hours) and analyzed by Western blotting.