The chromatin association of the FA core complex is impaired in FANCM-depleted cells. (A) EUFA867 lymphoblast and wild-type cells (GM02254A.L) were treated with MMC (150 ng/mL for 17 hours) and fractionated into S and P. α-tubulin and histone H3 are loading controls for S and P fractions, respectively. (B) HeLa cells transfected with the indicated siRNAs were analyzed by immunoblotting to insure FANCM or FANCA protein knockdown. (C) HeLa cells treated with the indicated siRNAs for 72 hours were treated with MMC (50 ng/mL for 17 hours). The whole-cell extracts were immunoblotted for the indicated proteins. The α-tubulin is a loading control for S fraction, and MCM3 was used as a loading control for P fraction. The crossreactive band are labeled with asterisks in panels A and C. (D) HeLa cells transfected with indicated siRNA for 72 hours were incubated without or with MMC (50 ng/mL for 17 hours) and then stained with anti-FANCG antibody, either without or with 0.3% Triton X-100 pre-extraction before fixation.