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. 2020 Apr 29;11:2094. doi: 10.1038/s41467-020-15783-y

Fig. 1. Phosphomimetic substitution at S429 enhances the E3 activity of the MDM2 homodimer.

Fig. 1

a Reduced SDS-PAGE showing autoubiquitination reactions catalyzed by GST-MDM2-419–C and its S429E substitution using fluorescently labeled Ub and visualized with an Odyssey CLx Imaging System. b Plot of relative ubiquitination activity corresponding to a. c Reduced SDS-PAGE showing autoubiquitination reactions catalyzed by GST-MDM2-419–C-His-MDMX-418–C and its MDM2-S429E substitution using fluorescently labeled Ub and visualized with an Odyssey CLx Imaging System. d Plot of relative ubiquitination activity corresponding to c. For b, d, data are presented as mean value ± SD from three independent experiments (n = 3). e Reduced SDS-PAGE showing autoubiquitination reactions catalyzed by GST-MDM2-419–C and its S429E substitution over the indicated times using fluorescently labeled Ub and visualized with an Odyssey CLx Imaging System. f A plot showing the rates of ubiquitination catalyzed by the forms of MDM2 assessed in e. The line represents the regression line from three independent experiments (n = 3). Asterisks in a, c, and e indicate non-reducible E1–Ub product. Uncropped gel images and InstantBlue-stained gels are shown in Supplementary Fig. 3.