p38α in hematopoietic cells protects mice from DSS-induced colitis. (A–C) 8-wk-old mice were given 3% DSS in drinking water for 6 days, followed by normal drinking water. The body weight change (A) and survival (B) of VavicreMapk14fl/fl (n = 5) and their littermates Mapk14fl/fl mice (n = 6) were measured. (C) At day 3 after DSS treatment, the colonic LPLs from wild type mice (n = 4) were isolated and analyzed by flow cytometry. Representative flow cytometry and mean fluorescence intensity (MFI) of p-p38 in Th (gating in CD45+CD90+CD3+CD4+), neutrophil (gating in CD45+Gr-1+CD11b+), DC (gating in CD45+MHCⅡ+CD11c+), macrophage (MΦ, gating in CD45+F4/80+CD11b+), and ILC3s (gating in CD45lowCD90high) were shown. FMO represents fluorescence minus one control. (D)Rorcgfp/+ reporter mice were exposed to 3% DSS for 3 days. Phosphorylated p38α in RORγt+TCRβ- ILC3s from colon was shown. Scale bars, 20 μm. (E) The isolated LPLs from the small intestine and colon were stimulated with different stimuli for 60 min in vitro and p-p38 in ILC3s (gating in CD45lowCD90high) was measured by flow cytometry. MFIs of p-p38 were shown. FMO represents fluorescence minus one control. (F) Representative flow cytometry and pooled analysis of IL-22 and GM-CSF production by ILC3s (gating in CD45+CD90+TCRβ−RORγt+). The colonic LPLs from naive VavicreMapk14fl/fl (n = 6) or Mapk14fl/fl (n = 6) littermate mice were in vitro–stimulated by PMA/ionomycin. (G) 8-wk-old VavicreMapk14fl/fl (n = 7) and their littermates Mapk14fl/fl (n = 4) were given 3% DSS in drinking water. At day 3, after DSS treatment, the colonic LPLs were ex vivo–stimulated by PMA/ionomycin and representative flow cytometry, and pooled analysis of IL-22 and GM-CSF production by ILC3s (gating in CD45+CD90+TCRβ−RORγt+) were shown. Data are representative of one of two independent experiments (A–E). Each symbol represents an individual mouse from two pooled experiments (F and G). Data are mean ± SEM and two-tailed Student’s unpaired t test (A, F, and G), log-rank (Mantel–Cox) test (B) were used for statistical analysis. *P < 0.05, **P < 0.01, ****P < 0.0001, ns, no significant difference.
Figure 1.

p38α in hematopoietic cells protects mice from DSS-induced colitis. (A–C) 8-wk-old mice were given 3% DSS in drinking water for 6 days, followed by normal drinking water. The body weight change (A) and survival (B) of VavicreMapk14fl/fl (n = 5) and their littermates Mapk14fl/fl mice (n = 6) were measured. (C) At day 3 after DSS treatment, the colonic LPLs from wild type mice (n = 4) were isolated and analyzed by flow cytometry. Representative flow cytometry and mean fluorescence intensity (MFI) of p-p38 in Th (gating in CD45+CD90+CD3+CD4+), neutrophil (gating in CD45+Gr-1+CD11b+), DC (gating in CD45+MHCⅡ+CD11c+), macrophage (MΦ, gating in CD45+F4/80+CD11b+), and ILC3s (gating in CD45lowCD90high) were shown. FMO represents fluorescence minus one control. (D)Rorcgfp/+ reporter mice were exposed to 3% DSS for 3 days. Phosphorylated p38α in RORγt+TCRβ- ILC3s from colon was shown. Scale bars, 20 μm. (E) The isolated LPLs from the small intestine and colon were stimulated with different stimuli for 60 min in vitro and p-p38 in ILC3s (gating in CD45lowCD90high) was measured by flow cytometry. MFIs of p-p38 were shown. FMO represents fluorescence minus one control. (F) Representative flow cytometry and pooled analysis of IL-22 and GM-CSF production by ILC3s (gating in CD45+CD90+TCRβ−RORγt+). The colonic LPLs from naive VavicreMapk14fl/fl (n = 6) or Mapk14fl/fl (n = 6) littermate mice were in vitro–stimulated by PMA/ionomycin. (G) 8-wk-old VavicreMapk14fl/fl (n = 7) and their littermates Mapk14fl/fl (n = 4) were given 3% DSS in drinking water. At day 3, after DSS treatment, the colonic LPLs were ex vivo–stimulated by PMA/ionomycin and representative flow cytometry, and pooled analysis of IL-22 and GM-CSF production by ILC3s (gating in CD45+CD90+TCRβ−RORγt+) were shown. Data are representative of one of two independent experiments (A–E). Each symbol represents an individual mouse from two pooled experiments (F and G). Data are mean ± SEM and two-tailed Student’s unpaired t test (A, F, and G), log-rank (Mantel–Cox) test (B) were used for statistical analysis. *P < 0.05, **P < 0.01, ****P < 0.0001, ns, no significant difference.

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