Figure 7. In a Mouse Model of Acute Colonic Inflammation, Members of the Silencing Complex Become Enriched at Promoter CpG Islands of Low Expression Genes.

(A) Mice were sham-inoculated (sham) or inoculated with ETBF. Two days post-inoculation, colon epithelial cells were extracted sequentially using cytoplasmic extraction buffer (Cyto), soluble nuclear buffer (Sol), and buffers with increasing NaCl concentration. Whole cell lysate was prepared separately (WCE). Blots from one set of representative mice are depicted. The value calculated for each fraction is the ratio of that fraction over the total of all fractions. The graphs represent the mean values for 3 separate mice +/− SEM. * p-value < 0.05 by one-tail t-test. (B) Co-immunoprecipitations for control IgG or anti-EZH2 antibodies were performed in nuclear lysates of colon epithelial cells from sham-inoculated mice (S) or ETBF-inoculated mice (E). Blots from one set of representative mice are depicted. (C) Using distal colon epithelial cells, ChIP was performed for IgG, EZH2, or DNMT1 and analyzed by quantitative PCR. The data presented is the mean of ChIP performed in samples from 3 sham and 3 ETBF mice +/− SEM. * p < 0.05 by one-sided t-test for the difference between the means. Values below the gene names are the expressed sequence tag (EST) counts for mouse intestine from the Unigene database. See also Figure S6.