Fig 2. RLIM promotes c-Myc ubiquitination.
(A) 293T cells were transfected with Flag-Ub, myc-c-Myc, HA-RLIM and HA-RLIMC596A in combinations as indicated. Ectopically expressed c-Myc was immunoprecipitated by anti-myc antibody followed by WB with anti-Flag antibody. (B) 293T cells were transfected as in (A) except using 6 × His-Ub instead of Flag-Ub. Ubiquitinated proteins were precipitated with nickel (Ni)-NTA beads and subjected to WB with anti-myc antibody. (C) 293T cells were transfected with His-Ub, HA-RLIM, myc-c-Myc and myc-c-MycDM (S62A and T58A) as indicated. Ubiquitinated proteins were precipitated with nickel (Ni)-NTA beads and subjected to WB with anti-myc antibody.