Figure S5.
Sustained IL-22 production by ILC3 in RBP-J KO mice is secondary to persistence of pathogens during the late phase of infection. WT and RBP-J KO mice were orally inoculated with 2 × 109 CFUs of C. rodentium. (A) qPCR analysis of Il17a in colon tissues (left) and LP mononuclear cells (right) at the indicated p.i. days. Data are pooled from two independent experiments; n ≥ 3 in each group. (B) qPCR analysis of Reg3g and Reg3b in colon tissues at the indicated p.i. days. Data are pooled from two independent experiments; n ≥ 3 in each group. (C) Periodic acid–Schiff staining in colons at day 14 p.i. (scale bars represent 20 µm). (D) Representative FACS plots (left) and cumulative data (right) of IL-22 production in LP CD4+ T cells (gated by CD3+CD4+) at day 14 p.i. Data are pooled from two independent experiments; n ≥ 3 in each group. (E) qPCR analysis of Reg3g and Reg3b in colon tissues at day 20 p.i. Data are pooled from two independent experiments; n = 3 in each group. (F) WT and RBP-J KO mice were orally inoculated with 2 × 109 CFUs of C. rodentium and were given neomycin sulfate individually at day 12 p.i. Representative FACS plots of IL-22–producing LP ILC3 (gated by CD45midCD3−Thy-1+) with or without neomycin treatment at day 14 p.i. (G and H) qPCR analysis of Il23a in colon tissues (G) and sorted CD64+ macrophages (H) at the indicated p.i. days. Data are pooled from two independent experiments; n = 3 in each group. (I) Representative FACS plots of LP neutrophils from WT and RBP-J KO mice with or without neomycin treatment at day 14 p.i. (J) qPCR analysis of Rbpj and Il1b in sorted LP neutrophils at day 14 p.i. Data are pooled from two independent experiments; n = 3 in each group. (K) Absolute numbers of neutrophils (CD11b+Ly6G+) and macrophages (CD11b+CD64+SiglecF−) in WT mice at day 14 p.i. Data are pooled from two independent experiments; n = 3 in each group. Data are shown as mean ± SEM; n.s., not significant; **, P < 0.01; two-tailed Student’s unpaired t test.