Two alanine-substituted FANCC mutations suppress IFN-γ–induced association of FANCC and STAT1 in B lymphocytes.

WCLs of JY cells stimulated with IFN-γ for 10 minutes (1 mg protein) were incubated with yeast-expressed GST-fusion proteins (10 μg) bound to glutathione-Sepharose beads. The glutathione-Sepharose affinity precipitates were analyzed by SDS-PAGE followed by immunoblotting. Upper panel: The Western blot was probed with antibodies to P-STAT1. Middle panel: The blot described in the top panel was stripped and reprobed with anti-STAT1 antibody. Lower panel: The same blot was reprobed with anti-FANCC to verify the input of the GST-fusion proteins. Lanes 1 and 2 were run as WCLs (60 μg of total proteins each) to show the effects of IFN-γ on expression and activation of STAT1.