Fig. 1. CircTmem241 knockout decreases ILC3 numbers.

a Top 10 highly expressed circRNAs were knocked down in CHILPs, followed by in vitro differentiation assay. ILC3s (CD3⁻CD19⁻CD127⁺CD45^lo RORγt⁺) were analyzed by FACS. n = 5 for each group. b CircTmem241 expression levels of indicated tissues were detected by Northern blot using biotin-labeled probes. 18 S RNA was used as a loading control. c Relative expression of circTmem241 was measured in indicated hematopoietic progenitors and mature cells by qPCR. Fold changes were normalized to endogenous 18 S. HSC (Lin⁻c-Kit⁺Sca-1⁺CD150⁺CD48⁻), MPP (Lin⁻c-Kit⁺Sca-1⁺CD150⁻CD48⁺), CLPs (Lin⁻CD127⁺c-Kit^intSca-1^intFlt3⁺α4β7⁻), CHILPs (Lin⁻CD25⁻CD127⁺Flt3⁻α4β7⁺Id2^GFP) and ILCPs (Lin⁻CD127⁺Flt3⁻c-Kit⁺α4β7⁺PLZF^GFP) were isolated from BM cells. ILC1s (CD3⁻CD19⁻CD45⁺CD127⁺NK1.1⁺NKp46⁺), ILC2s (Lin⁻CD127⁺CD90⁺KLRG1⁺Sca1⁺) and ILC3s (CD3⁻CD19⁻CD127⁺CD45^lo CD90^hi) were isolated from small intestinal lamina propria. T cells (CD3⁺CD45⁺) were isolated from thymus. B cells (NK1.1⁻CD19⁺CD45⁺) and NK cells (NK1.1⁺CD19⁻CD45⁺) were isolated from spleen. n = 3 biological independent experiments. d–f ILC1s (CD3⁻CD19⁻CD45⁺CD127⁺ gated), ILC2s (CD3⁻CD19⁻CD127⁺CD90⁺ gated), and ILC3s (CD3⁻CD19⁻CD45^low gated) were analyzed in small intestines of circTmem241^+/+ and circTmem241^−/− mice by FACS. n = 5 for each group. g Numbers of indicated ILCs in d–f were calculated. n = 5 for each group. h Immunofluorescence staining of ILC3s in small intestines from circTmem241^+/+ and circTmem241^−/− mice. Yellow arrows indicate ILC3s. Scale bar, 100 μm. i Lamina propria lymphocytes (LPLs) from circTmem241^+/+ and circTmem241^−/− mouse intestines were sorted and stimulated by IL-23 for 4 h, followed by IL-22⁺ ILC3 detection with FACS. n = 5 for each group. j IL-22 levels secreted by ILC3s from circTmem241^+/+ and circTmem241^−/− intestines were assayed by ELISA after stimulation by IL-23 for 24 h. n = 3 for each group. k Body weight changes of circTmem241^+/+ and circTmem241^−/− mice were analyzed after C. rodentium infection. n = 5 for each group. l Bacterial counts (CFUs) in feces of circTmem241^+/+ and circTmem241^−/− mice were measured after infection with C. rodentium for 6 days. n = 5 for each group. m Colon tissues from circTmem241^+/+ and circTmem241^−/− mice were analyzed by H&E staining. Scale bars, 100 μm. n Histopathological scores of colons from circTmem241^+/+ and circTmem241^−/− mice over C. rodentium infection. n = 5 for each group. **P < 0.01 and ***P < 0.001. Data were analyzed by an unpaired two-side Student’s t test and shown as means ± SD. Data are representative of at least three independent experiments. Source data are provided as a Source Data file.