title: GSM7308293: GFP+ ILC2, pre-transplant, rep2, snRNA-Seq; Mus musculus; RNA-Seq
library_construction_protocol: (in vitro) ILC2 + pcILC2: Lineage negative, ST2+ cells from the peritoneum and mesenteric lymph nodes were isolated and expanded in culture for 2 days in the presence of recombinant IL-7 and IL-33. ILC2s were expanded in IL-7 and IL-33 for 4 additional days, while the pcILC2 cells were treated with IL-12, IL-1b, and IL-18 (pcILC2). (ex vivo) GFP+ exILC2 post-BMT: Animals were euthanized with CO2 and the small intestine lamina propria (LP) was excised. LP lymphocytes were isolated using the Miltenyi LP dissociation kit (catalog 130-097-410) as per the manufacturer's instructions. Livers and lungs were digested in a solution of 1 mg/ml collagenase A (Roche) and 75 U DNase I (Sigma-Aldrich) in RPMI 1640 with 5% newborn calf serum. Digested tissues were treated with ACK lysis buffer to remove RBCs and were passed through 100 μm cell strainers. Leukocytes were collected at the interface of a 40%:80% Percoll (Sigma-Aldrich) gradient in RPMI 1640 with 5% NCS. The pelleted cells were washed in 1x DPBS with 2% FBS. Spleens and MLN were teased apart, treated with ACK lysis buffer, and washed in 1 x DPBS with 2% FBS. For all experiments, library preparation was performed according to the manufacturer's instructions (see Single Cell Chromium Next GEM Multiome ATAC+GEX, 10x Genomics, CG000338, Rev D). Nuclei were isolated from sorted GFP+ cells following the 'Low Cell Input Nuclei Isolation' protocol (CG000365, Rev C) where lysis buffer was used at 1x and lysis proceeded for 4 minutes. 10X Multiome