Immunodepletion and rescue. (A) Mitotic extracts were
immunodepleted with control IgG (lane 1), with a mixture of
anti-XCAP-C, -D2, -E, and -G (lanes 2–5), or with a mixture of
anti-XCAP-D2 and -G (lanes 6 and 7). Then a purified fraction of 13SC
(lane 3), 8SC (lane 4), 11SR (lanes 5 and 7), or buffer (lanes 2 and 6)
was added back into the depleted extracts. Aliquots of each extract
were analyzed by immunoblotting. (B) After incubating
sperm chromatin with each extract at 22°C for 2 h, chromatin was
isolated and bound proteins were analyzed by immunoblotting.
(C) Sperm chromatin was incubated with each extract
(1–7 as in A) at 22°C for 2 h, fixed, and
stained with 4′,6-diamidino-2-phenylindole. (Bar, 10 μm.)