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. 2021 Aug 4;14(6):1306–1322. doi: 10.1038/s41385-021-00431-5

Fig. 6. Ahr promotes Aiolos expression in ILC2s cell intrinsically.

Fig. 6

a LI and SI ILC2 commonly specific OCRs and differentially decreased peaks correlated genes were overlapped with LI and SI ILC2 common signature genes. b, c Purified LI ILC2s were treated with DMSO or FICZ (1 nM) for 24 h and analyzed with flow cytometry. Mean fluorescence intensity (MFI) of Aiolos (b) and PD-1 (c) gated on live cells were shown. d, f Expression of Aiolos and PD-1 in tissue ILC2s (Lin–GATA3+) from indicated organs of littermate wild-type or Ahr–/– mice was analyzed by flow cytometry. e, g Mixed bone marrow cells at 1:1 ratio from CD45.2-Ahr-deficient and CD45.1-wild-type mice were transferred to half-lethally irradiated Rag2–/–Il2rg–/– mice. Six weeks later, expression of Aiolos (e) and PD-1 (g) in ILC2s from indicated tissues was analyzed by flow cytometry. Percentages of Aiolos+ cells or PD-1+ cells gated on ILC2s from indicated origins (CD45.1+Lin–GATA3+ or CD45.2+Lin–GATA3+) are shown. Error bars are mean + SEM. Data were pooled from three to five mice of two independent experiments. h–j ILC2s were purified from the large intestine of wild-type (WT, dashed line) or Ahr–/– mouse and cultured in vitro in the presence of IL-7 and IL-33. Expression of IL-5 was analyzed by flow cytometry on day 3 (h) or day 10 (i) with brefeldin A added in the last 2 h. j Expression of Ki67 was analyzed by flow cytometry on day 10 after culture. k–o ILC2s were expanded for 5 days and infected with retrovirus expressing MIG or Ikzf3-MIG twice in the next 2 days. GFP+ virus-infected cells were purified and re-plated starting with equal cell numbers for another 48 h before cells were analyzed. k–n Expression of Aiolos (k), PD-1 (m), and Ki67 (n) gated on GFP+ cells was analyzed by flow cytometry. l MFI of Aiolos. o Absolute numbers of IL-5+ILC2s were total numbers of ILC2 time the percentages of IL-5+ cells. m–o Connected lines indicate cells from same mouse and same batch of experiment. b–o Data are from two to four independent experiments. d–g, l Error bars are mean + SEM. LI large intestine, SI small intestine.