FIGURE 6.

Effect of SUMOylation on the cleavage of eEF2. A, HeLa cells were transfected with SUMO1, SUMO2, or control vector and blotted with anti-C-terminal eEF2 antibody. Increased level of cleaved eEF2 was observed in the nuclear extracts of SUMO-transfected cells. HA blotting indicated the transfect efficiency of SUMO plasmids. Lamin A/C and GAPDH were used as nuclear and cytosolic markers, respectively. B, HeLa cells were transfected with siRNAs against SUMO1 and SUMO2/3, then fractionated after 48 h, blotted with indicated antibodies that showed consistency of decreased amounts of cleaved eEF2, and decreased the SUMOylation level. Data are representatives of three independent experiments. CE, cytosolic fraction; NE, nuclear fraction; WB, Western blot.