Figure 7. MAFF induces expansion of Clostridiales and regulates colonic homeostasis. Antibiotics-treated mice were inoculated with PBS, BTWT, or BTΔMAFF strains to generate SPF-Abx mice 4 wk before treatment with DSS. (A) Pooled Kaplan-Meier survival data of the five DSS colitis-sensitive experimental groups and matched PBS controls (n = 21–27 mice). All mice were treated with 4% DSS. (B) Blinded scoring of distal colon histology sections 10 d after initiation of DSS treatment (n = 7–9 mice). (C) DNA array was used to measure the gene expression profiles of PI−CD45−EpCAM+ colonic epithelial cells purified by FACS from the indicated SPF-Abx mice 4 wk after colonization. The differentially expressed genes (fold change > 2, P < 0.05, unpaired Student’s t test) were used to generate enriched pathways analyses, which were up-regulated in BTWT colonization (n = 4 mice) compared with BTΔMAFF strain (n = 4 mice). Full annotation is in Table S5. (D and E) The relative abundance of bacterial families identified with 16S rRNA analysis in feces (D; n = 16–21 mice per group) and RNA-Seq analysis of the Firmicutes gene expression in cecal samples of SPF-Abx mice collected at 4 wk after B. theta colonization (E), immediately before DSS treatment. The functional modules that aligned to the genome references of phylum Firmicutes and differentially expressed between BTWT (n = 4)- and BTΔMAFF (n = 3)-colonized mice are shown (log2 FoldChange>2, padj<0.05). Full annotation is in Table S4. (F) Concentrations of butyrate in cecal samples of SPF-Abx mice colonized with BTWT or BTΔMAFF strains measured by gas liquid chromatography (n = 3–4 mice for each group). (G) Gut microbial composition in cecal samples of BP mice, as in Fig. 3 A, was analyzed with 16S rRNA analysis and annotated with the taxonomic distribution at the family level. n = 5 mice for each group. Statistical analyses were performed with one-way ANOVA with Tukey’s multiple comparison test (B) or Student’s t test (F). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Error bars represent SEM.
Figure 7.

MAFF induces expansion of Clostridiales and regulates colonic homeostasis. Antibiotics-treated mice were inoculated with PBS, BTWT, or BTΔMAFF strains to generate SPF-Abx mice 4 wk before treatment with DSS. (A) Pooled Kaplan-Meier survival data of the five DSS colitis-sensitive experimental groups and matched PBS controls (n = 21–27 mice). All mice were treated with 4% DSS. (B) Blinded scoring of distal colon histology sections 10 d after initiation of DSS treatment (n = 7–9 mice). (C) DNA array was used to measure the gene expression profiles of PI−CD45−EpCAM+ colonic epithelial cells purified by FACS from the indicated SPF-Abx mice 4 wk after colonization. The differentially expressed genes (fold change > 2, P < 0.05, unpaired Student’s t test) were used to generate enriched pathways analyses, which were up-regulated in BTWT colonization (n = 4 mice) compared with BTΔMAFF strain (n = 4 mice). Full annotation is in Table S5. (D and E) The relative abundance of bacterial families identified with 16S rRNA analysis in feces (D; n = 16–21 mice per group) and RNA-Seq analysis of the Firmicutes gene expression in cecal samples of SPF-Abx mice collected at 4 wk after B. theta colonization (E), immediately before DSS treatment. The functional modules that aligned to the genome references of phylum Firmicutes and differentially expressed between BTWT (n = 4)- and BTΔMAFF (n = 3)-colonized mice are shown (log2 FoldChange>2, padj<0.05). Full annotation is in Table S4. (F) Concentrations of butyrate in cecal samples of SPF-Abx mice colonized with BTWT or BTΔMAFF strains measured by gas liquid chromatography (n = 3–4 mice for each group). (G) Gut microbial composition in cecal samples of BP mice, as in Fig. 3 A, was analyzed with 16S rRNA analysis and annotated with the taxonomic distribution at the family level. n = 5 mice for each group. Statistical analyses were performed with one-way ANOVA with Tukey’s multiple comparison test (B) or Student’s t test (F). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Error bars represent SEM.

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