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. 1998 Jan 6;95(1):132–137. doi: 10.1073/pnas.95.1.132

Figure 5.

Figure 5

Chromatin associates with native but not recombinant p55. (A) Coimmunoprecipitation of p55 with reconstituted chromatin. 100 μl of S-150 chromatin assembly extract alone (Ex) or chromatin (Ch) reconstituted with S-150 extract and hsp70 promoter-containing plasmid DNA (and purified by spin-column chromatography) were analyzed by immunoprecipitation with α-p55 and preimmune serum (Pre). Immunoprecipitates were analyzed for histones by SDS-PAGE and Western blotting using antibodies to histone H3. (B) p55 alone does not bind to nucleosomes. Baculovirus-expressed p55 immobilized on beads (20-μl bed volume) was incubated with 100 μl (∼20 mg/ml protein) of S-150 extract (Ex) and with hsp70 promoter-containing plasmid chromatin (Ch) reconstituted in the same extract and purified by spin-column chromatography. After washing in extract buffer, 0.1 M KCl, the pelleted beads and 10% of the supernatant (Sup) were boiled in 30 μl SDS-PAGE sample buffer. 10-μl samples were analyzed for histone H3 by Western blotting.