SRX20260871 GSM7308291_r1 SRP436558 PRJNA970390 SRS17591670 GSM7308291 GSM7308291 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA (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 NextSeq 2000 SRX20260872 GSM7308301_r1 SRP436558 PRJNA970390 SRS17591671 GSM7308301 GSM7308301 ATAC-seq GENOMIC SINGLE CELL other (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 NextSeq 2000 SRX20260873 GSM7308302_r1 SRP436558 PRJNA970390 SRS17591672 GSM7308302 GSM7308302 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA (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 NextSeq 2000 SRX20260874 GSM7308303_r1 SRP436558 PRJNA970390 SRS17591673 GSM7308303 GSM7308303 ATAC-seq GENOMIC SINGLE CELL other (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 NextSeq 2000 SRX20260875 GSM7308292_r1 SRP436558 PRJNA970390 SRS17591674 GSM7308292 GSM7308292 ATAC-seq GENOMIC SINGLE CELL other (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 NextSeq 2000 SRX20260876 GSM7308293_r1 SRP436558 PRJNA970390 SRS17591675 GSM7308293 GSM7308293 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA (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 NextSeq 2000 SRX20260877 GSM7308294_r1 SRP436558 PRJNA970390 SRS17591676 GSM7308294 GSM7308294 ATAC-seq GENOMIC SINGLE CELL other (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 NextSeq 2000 SRX20260878 GSM7308295_r1 SRP436558 PRJNA970390 SRS17591677 GSM7308295 GSM7308295 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA (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 Illumina NovaSeq 6000 SRX20260879 GSM7308296_r1 SRP436558 PRJNA970390 SRS17591678 GSM7308296 GSM7308296 ATAC-seq GENOMIC SINGLE CELL other (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 Illumina NovaSeq 6000 SRX20260880 GSM7308297_r1 SRP436558 PRJNA970390 SRS17591679 GSM7308297 GSM7308297 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA (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 Illumina NovaSeq 6000 SRX20260881 GSM7308298_r1 SRP436558 PRJNA970390 SRS17591680 GSM7308298 GSM7308298 ATAC-seq GENOMIC SINGLE CELL other (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 Illumina NovaSeq 6000 SRX20260882 GSM7308300_r1 SRP436558 PRJNA970390 SRS17591681 GSM7308300 GSM7308300 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA (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 NextSeq 2000