Fig. 2: CircKcnt2 knockout induces ILC3 activation and causes spontaneous colitis.

a mRNA levels of indicated genes were tested by qRT-PCR in ILC3s from DSS-treated circKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice. b Analysis of IL-17⁺ ILC3s (Lin⁻CD45⁺RORγt⁺ gated) in DSS-treated circKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice (***P < 0.0001). Numbers of total and IL-17⁺ ILC3s were calculated in the right panels. n = 5 for each group. c Geometric mean fluorescence intensity (MFI) of IL-17⁺ ILC3s was shown (***P = 0.0008). n = 3 independent samples. d ILC3s were isolated from DSS-treated mice and incubated with or without IL-23 for 24 h (**P = 0.0041). Secreted IL-17 was detected by ELISA. Veh vehicle. n = 3 independent samples. e Body weight changes were calculated. CircKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice were treated with 3% DSS for 7 days, followed by normal water for 7 days (***P = 0.0001). n = 5 for each group. f After DSS treatment, colon sections from circKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice were analyzed by H&E staining. Scale bar, 100 μm. g Colitis scores of indicated mice were analyzed (**P = 0.0071). n = 5 for each group. h After DSS treatment, survival rates of circKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice were monitored. n = 10 for each group. i CircKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice treated with 3% DSS for 5 days were administrated with FITC-dextran by gavage. Translocation of FITC-dextran into mouse serum was detected (***P = 0.0005). n = 5 independent samples. j IL-17⁺ ILC3 numbers were calculated in circKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice after DSS treatment (**P = 0.0035; ***P = 0.0008). n = 5 for each group. k H&E staining of colons from indicated mice treated with 3% DSS and administrated with anti-IL-17 or isotype IgG. Scale bar, 100 μm. CircKcnt2^+/+ and circKcnt2^−/− ILC3s (3 × 10⁴ cells per mouse) were transplanted into Rag1^−/−Il2rg^−/− mice. Two weeks later, mice were treated with DSS. l Colitis scores of indicated mice treated as in k were analyzed (**P = 0.0033). n = 5 for each group. m Body weight changes of mice in k were calculated (***P = 0.0005). n = 5 for each group. n CircKcnt2 expression was tested by qRT-PCR in ILC3s from H.h.-infected Il10^−/− mice (***P < 0.0001). n = 3 independent samples. o After H.h. infection, colon sections from circKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice were analyzed by H&E staining. Scale bar, 100 μm. p Colitis scores of indicated mice were analyzed. CircKcnt2^+/+Rag1^−/− and circKcnt2^−/−Rag1^−/− mice were infected with H.h. (**P = 0.0025). n = 5 independent samples. q Frequencies of fecal occult blood (FOB) in circKcnt2^+/+ and circKcnt2^−/− mice were analyzed (***P < 0.0001). n = 5 independent samples. r Body weights of circKcnt2^+/+ and circKcnt2^−/− mice were measured (**P = 0^.0065). n = 5 independent samples. s Colon lengths of circKcnt2^+/+ and circKcnt2^−/− mice were detected (*P = 0.0176⁾. n = 3 independent samples. t H&E staining of colon sections from circKcnt2^+/+ and circKcnt2^−/− mice. Scale bar^, 100 μm. u Colitis scores of circKcnt2^+/+ and circKcnt2^−/− mice (***P ⁼ 0.0007). n = 5 for each group. *P < 0.05, **P < 0.01 and ***P < 0.001. Data were analyzed by an unpaired 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.