Atac-seq of mus musculus: gut iILC2s
Gut ILC2s from C57BL/6 mice after A. muciniphila colonization were sorted, washed once with cold PBS, and lysed in cell lysis buffer (10 mM Tris-HCl pH 7.5, 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40, 0.1% Tween-20, 0.01% digitonin). After incubating on ice for 3 min, cells were washed once in wash buffer (10 mM Tris-HCl pH 7.5, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20), and centrifuge at 500 g for 10 min. Cell pellets were then incubated in transposition reaction buffer (10 mM Tris-HCl pH 7.6, 5 mM MgCl2, 10% dimethylformamide, 0.1% Tween-20, 0.01% digitonin, 33% 1 PBS) with 1.5 l pAG-Tn5 (15-1017, EpiCypher) at 37C for 30 min. DNA was purified using the QIAquick PCR Purification Kit (28104, Qiagen). Library PCR was performed using standard Illumina Nextera Dual Indexing primers. Libraries were sequenced using paired-end sequencing on Illumina NextSeq 500 platforms.