FIG. 7.

H3.3 and H2A.Z co-occupancy on the promoters of the heparanase and CD69 genes during transcriptional activation. (A) Schematic diagram indicating the experimental protocol used in the sequential ChIP assays. Note that the antibody order was reversed for all experiments and similar results were obtained (see Fig. S5 in the supplemental material). The primers used were located in the promoter regions of the CD69 and heparanase genes, as indicated. (B) CD69 and heparanase promoter DNA enrichment seen when chromatin was immunoprecipitated first with anti-H2A.Z antibody and then anti-H3 antibody. CD69 and heparanase data are shown for Jurkat T cells left NS or stimulated for 4 h with PI. Graphs show the n-fold changes in signal relative to the NS sample. Data pooled from two independent experiments are shown, with the mean ± the standard error plotted. (C) Sequential ChIP assays were performed as in panel B, except that anti-H3.3 antibody was used for secondary immunoprecipitation in place of anti-H3 antibody. Data are depicted as in panel B. Data pooled from two independent experiments are shown, with the mean ± the standard error plotted.