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. 2003 May 1;17(9):1141–1152. doi: 10.1101/gad.1070003

Figure 6.

Figure 6

(A) Tight association of GINS with chromatin. Sperm chromatin was incubated in egg extracts at 23°C for 15 or 30 min, and the extracts were then diluted with the chromatin isolation buffer containing 0 M, 0.4 M, and 0.8 M NaCl. The isolated chromatin fractions were resolved by SDS-PAGE and immunoblotted with the indicated antibodies. (B) Coimmunoprecipitation of GINS with other replication proteins from replicating chromatin. Sperm chromatin was incubated in egg extracts at 23°C for 45 min, and the isolated chromatin fractions were digested with micrococcal nuclease. The solublilized chromatin fractions were then immunoprecipitated with control, anti-Sld5, anti-Cdc45, and anti-Mcm2 antibodies. Immunoprecipitates were resolved by SDS-PAGE and immunoblotted. (*) A smeared band of cross-linked products of heavy and light chains of IgG eluted from the antibody-conjugated beads. In lane 5, only a small amount of Sld5 was extracted from anti-Sld5 antibody conjugated beads, and most Sld5 remained associated with the beads under the present extraction conditions.