Anti h3k27me3

Chromatin Immunoprecipitation Assay was performed on the IECs utilizing the commercially available EpiQuick High Sensitivity ChIp Kit P-2027 (Epigenetek, Farmingdale, NY), according to the manufacturer's instructions. Tissues were fixed with 1% formaldehyde for 10 min at 37 C, followed by glycine stop-fix solution. We used 4 μg of the following antibodies: anti-EZH2 (Millipore), anti-H3K27me3 (Epigentek), anti-phospho p65 (276) (Santa Cruz), anti-MUC1 Ab5 (Thermo Fisher Scientific).
The presence of IL-6 and TNF-α gene promoter sequences in immunoprecipitated DNA was identified by 34 cycles of PCR using the primer sequences published previously [34 (link)]. In control samples, primary antibody was replaced with non-immune IgG. All experiments were repeated at least three times.

Antibodies were purchased from the following suppliers: anti-H3K9me2, anti-H3K27me3 and anti-H3K9ac (EpiGentek, Farmingdale, NY); anti-H3K4me3, anti-H3K4me1 and anti-β-actin (Abcam, Cambridge, UK); negative rabbit IgG (Cell Signaling Technologies); anti-PRDM1 (sc-66015, Santa Cruz Biotechnologies, Dallas, TX); anti-TP53 (P6874 from Sigma and sc-98 from Santa Cruz); anti-mouse HRP-conjugated and anti-tubulin (Sigma); and anti-FOXA1 from Santa Cruz. Recombinant human TP53 protein (ref. 506165) was obtained from Calbiochem (San Diego, CA). The inhibitor 5-aza-2′-deoxycytidine (5-aza-dC) was purchased from Sigma-Aldrich and dissolved in DMSO.