Fig. 1.

FAAP20 is a FANCA-binding protein. (A) 293T cells stably expressed SFB-FANCA or SFB-FAAP20, respectively, were subjected to TAP and mass spectrometry analysis. Red indicates the bait protein and blue indicates the known or putative-associated proteins. Number of peptides recovered from mass spectrometry analysis is also presented. (B) 293T cells were transfected with constructs encoding SFB-FANCA along with constructs encoding Myc-FANCG or Myc-FAAP20. Coprecipitation and immunoblotting were carried out as indicated. (C) Lysates prepared from control or MMC-treated 293T cells were incubated with protein A agarose beads conjugated with rabbit IgG or anti-FAAP20 antibodies. Western blotting was performed using indicated antibodies. (D) Schematic illustration of wild-type and deletion mutants of FANCA used in this study. (E) 293T cells were transfected with constructs encoding SFB-FAAP20 along with constructs encoding Myc-tagged wild-type or deletion mutants of FAAP20. Precipitation and immunoblotting were conducted as indicated. (F) Pull-down assays were performed using bacterially expressed and purified GST-FAAP20 and MBP-fused FANCA (residues 674–1032) or FANCG. Immunoblotting were conducted using anti-MBP antibody.