MAD2L2 degradation in early anaphase is dependent on a destruction box and the proteasome. (A) Long time-course of MAD2L2 expression in U2OS after release from nocodazole with a representative cell cycle profile (of three repeats). (B) Stabilization of MAD2L2 levels by MG132 after release from double thymidine block. (C) In vivo ubiquitination of MAD2L2 on release from nocodazole. Left-hand three lanes: input blotted with anti-MAD2L2 (bottom) and anti-ubiquitin (top). Right-hand lanes: the anti-MAD2L2 immunoprecipitate blotted with anti-ubiquitin with IgG only control on the far right. (D) Stabilization of MAD2L2 by proteasome inhibition with MG132 after release from nocodazole. (E) The RXXL motif in vertebrate MAD2L2. Alignments of the first 19 amino acids of human (Hs), mouse (Mm), Xenopus (Xl), and chicken (Gg) MAD2L2. (F) MAD2L2 levels are stabilized by mutation of the D-box. Ectopically expressed myc-tagged MAD2L2 is degraded on release into G1 (left) and this is prevented by mutation of R6 and L9 to A (right).