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. 2002 Dec 1;16(23):3004–3016. doi: 10.1101/gad.249202

Figure 6.

Figure 6

Figure 6

Distinct roles of topo II and Plx1/aurora B in sister chromatid resolution. (A) Sperm chromatin was incubated with interphase LSS for 2 h, and then a half volume of CSF extract without (a,a′,a") or with (b,b′b") 10 μM ICRF-193 was added. After incubation for another 2 h, chromosomes were fixed and stained as in Figure 5A. Bar, 10 μm. (B) A kinetoplast DNA decatenation assay was used to measure topo II activity in CSF extracts in the absence (lanes 2–5) or presence (lanes 6–9) of 10 μM ICRF-193, or in mock-depleted (lanes 11–14) or Plx1/aurora B-depleted (lanes 15–18) CSF extracts. Samples were taken at the indicated times, and the DNA was deproteinized and run on a 0.8% agarose gel containing 0.5 mg/mL ethidium bromide. The substrate DNA was run in lanes 1 and 10. The catenated population was retained in the wells of the gel whereas the decatenated products (∼3 kb circular DNA, relaxed or supercoiled) migrated to the positions indicated. (C) Chromatin fractions were isolated from the interphase and mitotic assembly mixtures described in A and analyzed by immunoblotting with the indicated antibodies. A sample from a mock assembly reaction (without sperm chromatin) was analyzed in parallel (lane 1).