GSM7978069: GRIP1_IL4_rep3; Mus musculus; OTHER
CUT&RUN assay was performed using the EpiCypher CUTANATM ChIC/CUT&RUN kit v2.1 according to manufacturer's protocol. In brief, 106 cells were harvested for each condition, washed with 1X PBS, and resuspended in wash buffer, followed by incubation at RT for 10 min with activated Concanavalin A (ConA) beads. Cells were then permeabilized and incubated overnight on a nutator at 4°C with 1 μg of normal rabbit IgG (EpiCypher), KLF4 (proteintech, 1180-1-AP), or GRIP1 (Novus Biologicals, NB100-1756) antibody. Cell-bead antibody complex was then washed with cell permeabilization buffer and incubated with pAG-MNase (EpiCypher), followed by addition of 2 mM CaCl2 and incubation for 2 h at 4°C. Stop buffer was added to the reaction mixture and DNA fragments were released and purified with a CUTANA DNA purification kit (EpiCypher). DNA library construction was performed using the NEBNext Ultra II DNA Library Prep Kit from NEB (E7645L). The libraries were sequenced on Illumina NovaSeq 6000 system (50 bp paired-end) at the Genomics Resources Core Facility at Weill Cornell Medicine