Figure 2
From: Mdm2 enhances ligase activity of parkin and facilitates mitophagy
Mdm2 dose-dependently increases parkin self-ubiquitination in intact cells. HEK293A cells were transfected with HA-ubiquitin, myc- (A) or FLAG-parkin (B), and varying amounts of untagged Mdm2. Parkin was immunoprecipitated with anti-myc or anti-FLAG antibody, and its ubiquitination was determined by Western blot with anti-HA antibody. Note that Mdm2 progressively increases the ubiquitination of WT parkin. (C) Quantification of the level of parkin self-ubiquitination in the presence of different concentrations of Mdm2 from four independent experiments. Data are presented as means + S.E.M. ANCOVA analysis with Mdm2 concentration as a co-variate yielded significant effect of Mdm2 (p < 0.0001).
