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. 2013 Mar 15;288(18):12866–12879. doi: 10.1074/jbc.M112.413393

FIGURE 1.

FIGURE 1.

Accumulation of SUMO-modified Rta after MG132 treatment. A, 293T cells were transfected with pCR-SUMO-1 (lanes 1 and 6), pCR-SUMO-2 (lanes 2 and 7), or pCMV-R (lanes 3 and 8) or cotransfected with pCMV-R and pCR-SUMO-1 (lanes 4 and 9) or pCR-SUMO-2 (lanes 5 and 10) in the presence of 5 μm MG132 (lanes 6–10) or DMSO (lanes 1–5). At 24 h after transfection, the cells were washed with PBS containing 10 mm N-ethylmaleimide. B, 293T cells were transfected with pHA-SUMO-1 (lanes 1 and 6), pHA-SUMO-2 (lanes 2 and 7), pFLAG-Rta (lanes 3 and 8) or cotransfected with pFLAG-Rta and pHA-SUMO-1 (lanes 4 and 9) or pFLAG-Rta and pHA-SUMO-2 (lanes 5 and 10) and then treated with MG132 (lanes 6–10) or DMSO (lanes 1–5). In A, SUMO-Rta was immunoprecipitated (IP) using anti-FLAG antibody and detected by immunoblotting (IB) using anti-Rta antibody. In B, SUMO-Rta was immunoprecipitated using anti-FLAG antibody and detected using anti-HA antibody. Asterisks indicate bands detected nonspecifically. Rta* indicates where the Rta band is supposed to locate.