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. 2022 Jan 22;25(2):103790. doi: 10.1016/j.isci.2022.103790

Figure 4.

Figure 4

CPEB4 controls IL-22 protein production, by binding Il-22 mRNA

(A) Gene expression of Il-22, Il-17a, Il-17f, Il-1a, Il-1b, Tnfa, Ifnγ mRNA in colon extracts from untreated WT (n = 8) and CPEB4KO (n = 8) mice, relative to Gapdh. Mean ±SEM ∗∗∗∗p <0.0001 (two-way ANOVA test, multiple comparisons).

(B and C) Il-22 mRNA expression in CD4+ T cells (B) and ILC3 (LTi and NCR+) (C) cells from the colon lamina propria of WT and CPEB4KO mice (n = 3/group). Mean ±SEM ∗∗p = 0.0037; ∗LTi p = 0.0222; ∗NCR+p = 0.0472 (unpaired t-test).

(D and E) Frequency of CD4+ T cells(D) (WT, n = 11; CPEB4KO, n = 13) and ILC3 cells (E) (WT, n = 8; CPEB4KO, n = 9) in colon lamina propria. ∗∗p = 0.0042 (unpaired t-test).

(F) ELISA of IL-22 in CD4+ naive T cells isolated from WT (n = 7) and CPEB4KO (n = 7) mice treated for 72 h with IL-23 (50 ng/mL). Mean ±SEM ∗∗∗∗p < 0.0001 (unpaired t-test).

(G) ELISA of IL-22 from four 0.5-cm segments of colon from WT and CPEB4KO mice cultured ex vivo for 24 h. Mean ± SEM of six mice/genotype. ∗p = 0.026 (Mann-Whitney test).

(H) RT-qPCR of Saa1/2 and Ang4, mRNA from isolated intestinal epithelial cells from WT (n = 8) and CPEB4KO (n = 6) mice. Mean ±SEM ∗∗p = 0.0023, ∗p = 0.008 (Mann-Whitney test).

(I) Binding of CPEB4 to Il-22, Il-17a, Il-17f and Il-23r mRNA was measured by RNA-immunoprecipitation (RIP)-qPCR. Cpeb4 and Rpl0 mRNAs were used as positive and negative CPEB4 binding controls, respectively. IgG was used as IP control. Data are pooled from three biologically independent experiments for panels D, E, I or two for F, G, H.