Gr h300

Conventional ChIP was performed as previously described (Shi et al., 2003 (link)) using formaldehyde-crosslinked chromatin. Cells were treated with 100nM Dex for 2 h. Antibodies KDM1A/LSD1 (Abcam ab17721), GR-H300 (Santa Cruz sc-8992), H3K4me1 (Abcam ab8895), H3K4me2 (Millipore 07–030), H3K4me3 (Millipore 07–473), H3K9me2 (Abcam ab1220), H3K27ac (Millipore 17–683), and H3 (Abcam ab1791) were used. All histone modification antibodies have been validated by ENCODE for ChIP-seq (Egelhofer et al., 2011 (link)). ChIP-seq was performed on two biological replicates, each with 35 ± 4 million (mean ± SE) reads sequenced by the Illumina Genome Analyzer II (GA II). Quantitative PCR (qPCR) of independent ChIP samples was used for validation. ChIP-qPCR primer sets were as follows: BIRC3_TSS, 5′-GGTTATTACCGCTGGAGTTC-3′ and 5′-AAATGCGTCACCCAAATCC-3′; BIRC3_GRB, 5′-GATGGCCAGTAATGGAACTG-3′ and 5′-ATGCATCTCATCAGGGCATC-3′; CDKN1C_TSS, 5′-ACTAGTACTGGGAAGGTCC-3′ and 5′-TTCTTCTCGCTGTCCTCTC-3′; CDKN1C_GRB, 5′-AGGTCAGCTCACAGGATTG-3′ and 5′-CCCTTGCGCAAAGAGAAAG-3′; DUSP1_TSS, 5′-GTCAGACCACTTAACTGTGG-3′ and 5′-GCAAAGGCATGGAAGAGTAG-3′; DUSP1_GRB, 5′-CCAGGTGCATTACAGGTATC-3′ and 5′-CTTAGGCATGTGACCTTTGG-3′; PER1_TSS, 5′-CATCATGTTCTCTTGGCTGGTGG-3′ and 5′-AGGACGGCTGTCGTTTTGTTG-3′; PER1_GRE, 5′-CATCAGATTGGAAGTGGCAG-3′ and 5′-CGACCAGGTAGGCATCTC-3′.