Figure 4
From: Loss of ubiquitin E2 Ube2w rescues hypersensitivity of Rnf4 mutant cells to DNA damage
Increased and prolonged FancD2/I monoubiquitination induced by MMC in ΔRNF4 is rescued by UBE2W inactivation.
Wild type cells and cells deficient for ΔRNF4, ΔUBE2W and ΔRNF4 ΔUBE2W were treated with MMC (100ng/ml) for 1H. (a) Proteins were analysed by western blot using antibodies against FancI (top panel) and FancD2 (bottom panel). The recovery time after MMC treatment of each protein extraction is indicated on each lane of the top panel in hours. Non-ubiquitinated and mono-ubiquitinated form of FancI and FancD2 are indicated by an arrows head on the left inside (respectively: I>, D2> and I-Ub>; D2-Ub>). Asterisk indicates non specific band (*). Molecular weight markers are indicated on the right inside (kDa). (b,c) quantification of the ratio of ubiquitinated protein to total protein has been determined by three independent LiCOR experiment for FancI and FancD2. LiCOR images can be seen in supplementary Fig. S3.
