CUT&Tag of mus musculus: adult female esophagus
CUT&Tag assay and DNA library construction for mouse esophageal tissues were conducted by SeqHealth (Wuhan, China) using the Hyperactive Universal CUT&Tag kit (Vazyme, China). Tissues were treated with collagenase and DNase I at 37°C for 2 h, then nuclei were isolated by centrifugation at 2000 g, 4°C for 10 min. Approximately 50,000 nuclei were bound to magnetic beads and incubated with H3K27ac antibody (abcam Cat# ab4729, USA) overnight at 4°C. After secondary antibody incubation, pA/G-Tnp was added, followed by TTBL for DNA fragmentation. DNA was then extracted, amplified, purified, quality-checked, and sequenced on the Illumina NovaSeq 6000 PE150 platform.