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Fig. 4. Reticulocyte lysate contains a Cdh1-dependent destruction activity. (A) Destruction of Aur-A in reticulocyte lysate was analysed as follows. Following coupled in vitro transcription-translation of the indicated templates for 90 minutes in rabbit reticulocyte lysate, 100 µg ml-1 cycloheximide (CHX) was added to inhibit further protein synthesis. Reaction mixes were shifted to 23°C and samples were taken at subsequent times for analysis by SDS-PAGE followed by autoradiography. Five minutes prior to cycloheximide (CHX) addition, 10 ng µl-1 Cdh1(1-125) protein was added as indicated. (B) Xenopus egg CSF extract was incubated for 5 minutes with Cdh1 protein, reticulocyte lysate (RL), or reticulocyte lysate and Cdh1(1-125) protein. Calcium was added to trigger exit from meiosis I. Samples were taken at the indicated times and analysed by PAGE and western blot for endogenous Xenopus Aur-A.





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