Fig. 3

Evi5 interacts with c-Myc to prevent its degradation. a TU212 cells stable expressing of either Flag-Evi5 or vector control (VC) were treated with 10 μM CHX for the indicated time. The whole cell lysate were detected by western blot and with the quantification plot based on scanning densitometry analysis. b Control or Evi5 KO TU212 cells were treated with 10 μM CHX for the indicated time. The whole cell lysate was detected by western blot and with the quantification plot based on scanning densitometry analysis. c TU212 cells transfected with indicated plasmids for 36 h were treated with MG132 during the last 4 h before lysis. Lysates were subjected to pulldown with TUBE2 resin to enrich ubiqutinated proteins, followed by western blot with anti-c-Myc antibody. d TU212 cells transfected with indicated plasmids for 36 h, were subjected to Flag M2 beads purification. Bound proteins were analyzed by western blot as indicated. e Lysates from TU212 cells were immunoprecipitated with either an antibody against c-Myc or a nonspecific IgG and analyzed by western blot as indicated. f Lysates from TU212 cells were immunoprecipitated with either an antibody against Evi5 or a nonspecific IgG and analyzed by western blot as indicated