Figure 3

Wwox-deficient cells exhibit increased homologous recombination, which is responsible for enhanced IR survival. In bar charts, Wwox-deficient cells are depicted as black and Wwox-sufficient cells are depicted as white. Western blot (WBs) below bar charts demonstrate transfection efficiency and Wwox expression. (a) Bar graph indicating % of cells positive for GFP as indication of relative HR, with corresponding immunoblots of U87 cells carrying HR reporter, DR-GFP, and transfected with siCtrl, siWwox and siWwox+myc-Wwox plasmid resistant to silencing. (b) Graph indicating % GFP-pos HeLa cells DR-GFP transfected with empty vector or Myc-Wwox. (c) Bar graph indicating relative mRNA as a measure of C-NHEJ in H1299 cells carrying pHW1 and transfected with siCtrl and siWwox. (d) Bar graph depicting fraction of GFP-pos cells in H1299 EJ2 cells indicating relative Alt-NHEJ efficiencies following transfection with siCtrl, siWwox+myc-Wwox and siWwox. (e) Bar graph indicating % of GFP-pos cells as a measure of SSA in HeLA Sa26 cells following transfection with empty vector and myc-Wwox. (f) Schematic of the four DSB repair pathways: NHEJ, HDR, SSA and Alt-NHEJ and corresponding changes in efficiency dependent on Wwox expression (right). (g) Line graph indicating % survival of MEFs with and without Rad51 silencing (denoted as R51 in embedded WBs) following exposure to designated IR doses. Solid lines indicate survival of MEFs with normal Rad51 expression, whereas dashed lines indicate Rad51-silenced cells. (h) Line graph depicting % survival of 231/Wwox cells with and without doxycycline following exposure to various levels of cisplatin for 2 h. (i) Line graph depicting % survival of MEFs following exposure to various levels of cisplatin for 2 h. (a–e and g–i) Data from three independent experiments were subjected to two-tailed Student’s t-test.