GSM6754115: Hypoxia_HIF1A_NB100-134_2; Homo sapiens; OTHER
In brief, 105 of the 3D organoid cells were used for the CUT&Tag. The experiments were based on digitonin (Sigma, D5628) mediated cell permeabilization and concanavalin A coated magnetic beads (Bangs Laboratories, BP531) immobilization. Two biological replicates were used for all experiments. H3K27ac (Abcam, ab4729), HIF1a(Novus Biologicals,NB100-134) and BACH1(R&D, AF5776) were used as the primary antibodies. Anti-goat (Sigma, SAB3700280) and anti-rabbit (EpiCypher, 13-0047) secondary antibodies were followed for 1-hour incubation. PA-Tn5 (EpiCypher, 15-1117) was loaded for 1-hour incubation. The cleaved DNA by Tn5 was extracted using a DNA Clean & Concentrator-5 Kit (Zymo Research, D4013). Library preparation was conducted using IDT primers (Illumina, 20027213) and NEB PCR enzyme mix (NEB, M0541S).