title: GSM4041766: ATAC_IL33_CGRP_2; Mus musculus; ATAC-seq
library_construction_protocol: Lungs were digested with the lung dissociation kit (Miltenyi Biotec) and subsequently enriched for CD90.2+ cells using CD90.2 MicroBeads (Miltenyi Biotec). Lung ILCs were isolated by fluorescence-activated cell sorting as 7AAD- CD45+ CD90.2+ CD127+ Lineage (CD3, CD4, CD8, CD11b, CD11c, CD19, NK1.1, TCRβ, TCRγδ)- cells. 6,000 viable ILC2s were sorted into DPBS supplemented with 2% FCS. Cells were pelleted by centrifugation and stored in BambankerTM cell freezing media (LYMPHOTEC Inc.) at -80 °C. For ATAC-seq library preparation, cells were thawed at 37 °C, washed once with PBS, and lysed and tagmented in 1X TD Buffer, 0.2ul TDE1 (Illumina), 0.01% digitonin, and 0.3X PBS in 40ul reaction volume following the protocol described in Corces at al. (Corces et al., 2016). Transposition was performed at 37 °C for 30 min at 300 rpm. The DNA was purified immediately with the MinElute PCR purification kit (Qiagen). The complete eluate was then amplified with PCR, as follows. First, 5 cycles of pre-amplification were performed using indexed primers with NEBNext High-Fidelity 2X PCR Master Mix (NEB). The number of additional cycles was assessed by SYBR Green quantitative PCR. After purifying the final library with the MinElute PCR purification kit (Qiagen), the library was quantified with the Kapa Library Quantification Kit (Kapa Biosystems) and a Qubit dsDNA HS Assay kit (Invitrogen). Libraries were sequenced on an Illumina NextSeq 550 system with paired-end reads of 37 base pairs in length.