Fig. 7: OLFM4 promotes RORγt and IL-22 expression through ASK1-p38 MAPK pathway.
From: OLFM4 modulates intestinal inflammation by promoting IL-22+ILC3 in the gut
A MFI of p-p38-Thr180/Tyr182 and (B) p-ASK1-Ser967 in ILC3 from C. rodentium infected mice was analyzed by flow cytometry (n = 3). C p38 phosphorylation level (Thr180/Tyr182) in MNK3 cells after transfection with si-OLFM4. D Expression level of RORγt in indicated MNK3 cells (n = 3). E The secretion level of IL-22 in the culture supernatant stimulated (IL-1β, IL-23, and IL-7) for 48 h (n = 4). F mRNA levels of Il23r, Il22, Ccr6, and Ncr1 in stimulated indicated MNK3 cells (n = 3). G Western blotting of the phosphorylation level of p38 in MNK3 cells after transfected with plasmids encoding OLFM4 (OEOLFM4) and empty vector (OEEV). H RORγt expression level in stimulated MNK3 cells after overexpression of OLFM4 (n = 4). I Secretion level of IL-22 in culture supernatant (n = 4). J mRNA levels of Il23r, Il22, Ccr6, and Ncr1 in stimulated indicated MNK3 cells (n = 4). Expression levels of (K) RORγt and (L) IL-22 in stimulated indicated MNK3 cells after treated with SB203580 or DMSO (n = 4). M Secretion level of IL-22 in the culture supernatant (n = 4). N Western blotting of ASK1 phosphorylation level in MNK3 cells after overexpression of OLFM4. Expression levels of (O) p-p38-Thr180/Tyr182 and (P) RORγt in stimulated MNK3 cells after overexpression of OLFM4 and treated with NQDI-1 (n = 3). Q Secretion level of IL-22 in the culture supernatant of indicated MNK3 cells (n = 3). Data are presented as mean ± SEM, and statistical significance was determined using two-tailed unpaired Student’s t test (A, B, D–F, H–J), or two-way ANOVA (K–M, O–Q). ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001. The experiments were performed three times (at least three replicates).
