Anti stat4 sc486

ChIP-Seq was performed using at least 10 million sorted NK cells without or with cytokine stimulation. After chemically cross-linking cells, chromatin was fragmented by sonication and immunoprecipitated by anti-H3K27Ac (ab4729; Abcam), anti-H3K4me3 (04–745; Millipore), anti-H3K27me3 (07–449; Millipore), anti-H3K4me1 (ab8895; AbCam), anti-p300 (sc585; Santa Cruz), anti-STAT4 (sc486; Santa Cruz), anti-STAT5 (ab7969; AbCam) or anti-T-bet (sc21003; Santa Cruz). After recovering purified DNA, 10 ng or more of DNA was used to generate libraries according to the vendor’s manual for the Illumina platform (E6240S/L; New England BioLabs). Illumina HiSeq 2500 or Genome Analyzer II was used for 50-cycle single-read sequencing.

Recombinant human IL-2 was obtained from the NCI repository. Recombinant murine IL-4, IL-12 (p70), and IL-13 were purchased from Peprotech (Rocky Hill, NJ, USA). Purified Lipopolysaccharide (LPS) in lyophilized powder from Escherichia coli 0111:B4 was purchased from Sigma (St. Louis, MO, USA). Anti-STAT6 (Sc-981) and anti-STAT4 (Sc-486) monoclonal antibodies were obtained from Santa-Cruz Biotechnology (Santa Cruz, CA, USA) and anti-STAT5 was from R&D Systems (Minneapolis, MN, USA). Anti-acetylated histone H3 (AcH3; 06-599B) and anti-histone H3 lysine 27 trimethylation (H3K27me3; 07-449) were obtained from Upstate Biotechnology (Millipore, Billerica, MA, USA).