ChIP assays were performed as described previously with minor modifications12 (link). Briefly, purified rat SCs (~20 million cells) grown in the proliferation or differentiation (9 h in 1 mM cAMP-containing medium) condition were fixed for 10 min at room temperature with 1% formaldehyde–containing medium. Nuclei were isolated and sonicated in sonication buffer (10 mM Tris-HCl, pH 8.0; 1 mM EDTA; 0.5 mM EGTA; and protease inhibitor cocktail). Sonicated chromatin (~300 μg) was used for immunoprecipitation via incubation with antibodies overnight at 4 °C. Prerinsed protein A/G PLUS-agarose beads (50 μl) were added to each ChIP reaction and incubated for 1 h at 4 °C. The beads were then incubated in 200 μl elution buffer at 65 °C for 20 min to elute immunoprecipitated materials. The ChIP–seq libraries were prepared using the NEBNext ChIP-seq Library Prep Master Mix Set for Illumina (NEB, E6240L) and then were run on the Illumina sequencer HiSeq 2000. Two ChIP–seq replicates were performed for diff_HDAC3, p300, and IgG, and one replicate was performed for Pro_HDAC3, siHdac3_ H3K27ac, Scr_H3K27ac, H3k27ac, and H3K4me1. Antibodies against the following proteins were used: HDAC3 (Santa Cruz Biotechnology, sc-11417), p300 (rabbit; Santa Cruz, sc-585), H3K27ac (rabbit; Abcam, ab4729), and H3K4me1 (rabbit; Abcam, ab8895).