Fig. 1. FA family proteins FANCD2 and FANCI are significantly reduced in MTDH knockout mice and MTDH-depleted cancer cells and increased in MTDH overexpressed cancer cells.

(A) Expression of FANCD2 and FANCI protein was detected and quantified by Western blotting in the brain (Br), liver (Li) and spleen (Sp) from wild type mice and two MTDH homozygous knockout mice (MTDH KO1 and MTDH KO2). Rad51 and β-actin were also analyzed by Western blotting as controls for other DNA repair proteins and loading. (B) Quantification of Western blots in (A). With the exception of Rad51, all data are relative to wide type brain. For Rad51 data are relative to MTDH^−/− 2 spleen. (C, D) FANCD2, FANCI and MTDH were detected and quantified by Western blotting in parental Hec50 cells and MTDH CRISPR knockout Hec50 cells in the absence or presence of DNA damage via cisplatin for 24 h. Labels denote ubiquitinated (Ub) FANCI and the long (L) and short (S) isoforms of FANCD2. β-actin was used as a loading control. (E,F) FANCD2, FANCI and MTDH were detected and quantified by Western blotting in empty vector transfected Hec50 cells and MTDH overexpressed Hec50 cells. β-actin was used as a loading control. Each figure is representative of three independent experiments.