Fig. 4.
Eco2 is a substrate of Cdh1-activated APC. (A) Radiolabeled in vitro-translated proteins (indicated at Left: cyclin B, sororin, xEco2, xEco1, and xEco2KEN) were incubated in interphase extract (IE), extract supplemented with cyclin B (+CycB), or supplemented with Cdh1 protein (+Cdh1). Aliquots were removed at the indicated times (t = 0, 45, 90, and 135′), separated by SDS/PAGE, and the gel was dried and exposed in a phosphor storage cassette. The phosphorimages are shown. Loading controls are shown in Fig. S6. (B) APC-dependent degradation of endogenous Eco2 protein interphase extract was analyzed by immunoblot for Smc3, Eco2, and sororin at the indicated times following the addition of buffer alone (IE), recombinant cyclin B (+CycB), or Cdh1 (+Cdh1).