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Enhancement of SUMO modification of dominant-negative Drp1. HEK-293 cells were transiently transfected with pcDNA3-V5-Ubc9, pcDNA3-HA-SUMO3, and/or pcDNA3-V5-His-Drp1 WT or pcDNA3-V5-His-Drp1 K38A, and CS2+empty vector. Protein extracts were obtained in the presence of 20 mM NEM, processed as in Fig. 2A, and analyzed by immunoblotting. Arrowheads indicate modified doublet; asterisk indicates unmodified Drp1.
