Adult differentiated jejunal epithelial cells; middle 1/3 of intestine
genotype
Cdx2f/f; Villin-cre
protocol
Villi from middle 1/3rd (jejunum) of mouse small intestine were isolated by incubating in 5mM EDTA in PBS for 45 minutes at 4°C. Cells were cross-linked and immediately snap-frozen in liquid nitrogen for subsequent ChIP analysis.
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Whole cell extracts from villi fixed with 1% formaldehyde were sonicated and incubated overnight with Dyna beads (Thermo Fisher Scientific) conjugated with 5 ug KDM6A antibody (Cell Signaling Technology, 33510) for immunoprecipitation. ThruPLEX DNA-seq Kit (Rubicon Genomics) was used to prepare libraries, and 75 bp single-end reads were sequenced on an Illumina NextSeq 500 instrument.