Fig. 1. Various sensitivities of FA knockout mutants identify the FA core complex catalytic module.

(A) Depiction of three protein interaction modules within the FA core complex and the FANCM-FAAP24 complex. See also Fig. S1A-C.

(B) Clonogenic survival of wild-type HCT116 (WT), FANCB⁻, FANCG^−/−, and FANCB⁻ G^−/− mutants treated with mitomycin C. See also Fig. S1D-E and S2.

(C) Clonogenic survival of wild-type HCT116 (WT), FANCB⁻, FAAP20^−/−, and FANCB⁻ FAAP20^−/− mutants treated with mitomycin C. See also Fig. S2.

(D) Clonogenic survival of wild-type HCT116 (WT), FAAP20^−/−, FANCG^−/−, FANCB⁻, and FANCL^−/− mutants treated with mitomycin C.

(E) Immunoblots detecting MMC- or cisplatin-induced monoubiquitination of chromatin-bound FANCD2 and FANCI in wild-type HCT116 cells (WT) and the indicated knockout mutants. See also Fig. S3A-B.

(F) Immunoblot detecting MMC- or cisplatin-induced monoubiquitination of chromatin-bound FANCD2 in wild-type chicken DT40 (WT) and indicated knockout mutants. *FANCG gene localizes in the single Z chromosome and only one allele exists in DT40 cells. See also Fig. S3C-D.

(G) Immunoblots detecting FANCL auto ubiquitination in protein extracts prepared from 293T cells stably expressing SPB-FAAP100 and/or SFB-FANCL with or without MMC treatment.. Protein extracts from indicated cell lines were subjected to S beads pull-down and immunoblotted by anti-ubiquitin or Flag antibodies.

(H) Superose 6 gel filtration profiling of the FA core complex in HCT116 WT, FANCG^−/−, and FANCL^−/− mutants. Nuclear extracts were fractionated by Superose 6 gel filtration and the indicated fractions were immunoblotted (IB:) with FANCL or FAAP100 antibodies. The arrow indicates the elution position of the FA core complex (670kD). Fraction 21 marks the void.

Error bars in survival curves were derived from SDs from 4–6 independent experiments with triplicates.