GSM4675977: R204X_plusDox_H3K27ac_1070_1316; Mus musculus; ChIP-Seq
ChIP was performed as previously described (Obier et al, 2016) with the following modifications. For H3K27ac only KIT+ Progs were crosslinked only in 1% formaldehyde (single crosslinking, as opposed to double crosslinking for RUNX1). For single crosslinked cells nuclei were sonicated for 4 cycles of 30s on/30s off using a Picoruptor (Diagenode). Immunoprecipitation was carried out overnight at 4°C using 2 µg of RUNX1 antibody (ab23980, Abcam), or for four hours at 4°C using 2 µg of H3K27ac antibody (??) coupled to 15 µg Dynabeads Protein G (Invitrogen) per 2 x 106 cells. DNA from 2-3 immunoprecipitations was pooled for RUNX1, but just 1 immunoprecipitation for H3K27ac, and extracted using Ampure beads (Beckman Coulter). ChIP libraries were generated using the KAPA hyper prep kit, libraries were size selected to obtain fragments between 150 and 450 bp