XB-IMG-119220
Xenbase Image ID: 119220
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Figure 3. Topoisomerase II is modified by SUMO-2/3 during mitosis in unperturbed cycling extracts. Xenopus cycling egg extracts were supplemented with 2,000 sperm nuclei/μl and incubated at 23°C. Samples were taken from extracts at the indicated times. Aliquots of each sample were directly subjected to SDS-PAGE and Western blotting analysis to determine cyclin B abundance (fourth panel) and phosphorylated ERK1 levels (bottom). Chromatin was separately prepared from samples taken at each time point, as described in Materials and methods. The chromatin preparations were analyzed by Western blotting with antibodies against topoisomerase II (top), SUMO-2/3 (second panel), and SUMO-1 (middle). Aliquots were also removed at the indicated times and analyzed for DNA morphology after addition of Hoechst 33342 DNA dye (not depicted). We observed condensed metaphase chromosomes at 75 min, and anaphase figures at 85 min after the start of the reaction. Image published in: Azuma Y et al. (2003) Copyright © 2003, The Rockefeller University Press. Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View |