Immunoprecipitation and immunoblotting was carried out as described previously (40 (link)). Briefly, cells were harvested and lysed in NTEN buffer (0.5% NP40, 50 mM Tris–HCl pH 8.0, 2 mM EDTA, 150 mM NaCl and 10% glycerol) supplemented with a complete protease inhibitor cocktail (Roche). For the immunoprecipitation assays, 1mg protein from the cell lysates was incubated with anti-HA (Roche) antibody and separated using protein G beads. The proteins were resolved on a 6% SDS-PAGE gel and transferred onto PVDF membrane (Millipore). The membranes were blocked using 5% skimmed milk in TBST buffer (50 mM Tris–HCl pH 8.0, 150 mM NaCl, and 0.1% Tween-20) and incubated with primary antibody for 12 h at 4°C. The primary antibodies against FANCM (1:50, sc-101389), BRCA-1 (1:100, sc-6954), MCM3 (1: 500, sc-365616) that were used for western blot analysis were purchased from Santa Cruz. Anti-MRE11 (1:2000, 611366) antibodies were obtained from BD Biosciences; and the rabbit polyclonal antibody for FANCJ (1:1000, ab49657) was obtained from Abcam. Anti-HA antibody (1:2000, 12CA5) was obtained from Roche. The membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies, developed by chemiluminescence, and imaged using Chemidoc (GE healthcare LAS 4000).