α h3k27ac

α-H3K27ac (abcam ab4729) was used for ChIP-seq using on 10 million Kasumi-1 AML1-ETO-FKBP cells at 0- and 24-hr post-treatment with dTAG-47 and with Drosophila S2 cell spike-in. Cells were cross-linked with 1% formaldehyde for 10 minutes RT and quenched with 125 mM Glycine. Following cell lysis, chromatin was sonicated with a Biorupter (Diagenode) to generate 300–600 bp chromatin fragments and immunoprecipitated with antibody plus Protein A:G beads. Library construction was carried out using the NEBNext Ultra II DNA Library Prep Kit and sequenced on the Illumina NovaSeq (PE-100) at Vanderbilt University Medical Center VANTAGE core. Data Analysis: Adaptors were trimmed with Trimmomatic-0.32 prior to aligning to the hg19 genome with Bowtie2 (v 2.2.2). Spike-in reads were removed and quantified. Bigwig files were generated using deepTools and normalized based on spike-in reads.

ChIP-seq was performed as described previously (23 ). For each assay, 4 – 10 μg antibodies were used: α-NEUROD1, Cell Signaling Technology, #4373, RRID:AB_10549071; α-BRD2, Cell Signaling Technology, #5848, RRID:AB_10835146; α-BRD3, Bethyl, A302-368A, RRID:AB_1907251; α-BRD4, Bethyl, A700-004, RRID:AB_2631885; α-H3K27Ac, Abcam, ab4729, RRID:AB_2118291. The ChIP-seq analysis was detailed in supplemental methods.