library_construction_protocol: CUT&RUN was performed using the Epicypher CUTANA ChIC/CUT&RUN Kit (Epicypher, #14-1048). Briefly, 48 hours post-transfection, 500k HEK293T cells were detached and harvested using 0.5mM EDTA (Fisher, #BP2482-500), washed once with 1X PBS and then resuspended in 300ul of wash buffer. 100ul aliquots (~1/3 of each 24 well) of cells were processed for H3K27ac antibody (Epicypher, #13-0045). Cells were first immobilized on concanavalin A beads, and then incubated with respective antibody (0.5ug/sample) overnight at 4°C in antibody dilution buffer (cell permeabilization buffer + EDTA). On the following day, cells were washed twice with cell permeabilization buffer. After washing the beads, pAG-MNase was added to the immobilized cells and then incubated for 2 hours at 4°C to digest and release DNA. Libraries were prepared using the CUT&RUN Library Prep Kit (Epicypher, #14-1001) and DNA was quantified using the Qubit HS dsDNA High Sensitivity Kit (Thermo Fisher) and sequenced at a depth of 15M paired end reads per sample on a NextSeq 2000 (High Output 2x100). Samples were normalized using an E. Coli spike-in DNA (Epicypher, #18-1401).