Figure 3. Non-specific 7-6IgA alters gene expression profile of B. theta in vivo. (A) Experimental scheme. Rag1−/− mice were treated with metronidazole and ciprofloxacin and colonized with chloramphenicol-resistant B. theta upon concomitant transplantation of 7-6IgA–secreting hybridoma cells, or control parental P3U1 myeloma cells under the dorsal skin. BP, backpack. (B) ELISA measurements of the total and anti-OVA IgA in fecal samples collected from BP 7-6IgA mice, as in A, and age-matched C57BL6 WT mice (n = 5 mice for each group). (C) The z-score of B. theta PUL genes from RNA-Seq of the colonic content of BP P3U1 and BP 7-6IgA mice (n = 4 mice for each group). A full set of log2 FoldChange≥1 and padj≤0.05 genes is shown in Table S2. The values of variance-stabilizing transformations were used to generate the heat map. (D) Schematic diagram of the MAFF genes and flanking regions in the indicated Bacteroidales strains. (E) The expressions of BT2268 and its orthologues were measured by qPCR in indicated Bacteroidales strains. Large intestinal content (Li-Cont) and mucus (Li-Muc) were collected 4 wk after colonization of SPF WT mice previously treated with antibiotics (SPF-Abx mice; n = 3–11 mice). The relative values of Li-Muc in comparison with Li-Cont are plotted in the graph. Each plot in B and E and each column in C represents the result obtained from independent mice. Statistical analyses were performed with Student’s t test in B and E. *P < 0.05 and ***P < 0.001. Error bars represent SEM.
Figure 3.

Non-specific 7-6IgA alters gene expression profile of B. theta in vivo. (A) Experimental scheme. Rag1−/− mice were treated with metronidazole and ciprofloxacin and colonized with chloramphenicol-resistant B. theta upon concomitant transplantation of 7-6IgA–secreting hybridoma cells, or control parental P3U1 myeloma cells under the dorsal skin. BP, backpack. (B) ELISA measurements of the total and anti-OVA IgA in fecal samples collected from BP 7-6IgA mice, as in A, and age-matched C57BL6 WT mice (n = 5 mice for each group). (C) The z-score of B. theta PUL genes from RNA-Seq of the colonic content of BP P3U1 and BP 7-6IgA mice (n = 4 mice for each group). A full set of log2 FoldChange≥1 and padj≤0.05 genes is shown in Table S2. The values of variance-stabilizing transformations were used to generate the heat map. (D) Schematic diagram of the MAFF genes and flanking regions in the indicated Bacteroidales strains. (E) The expressions of BT2268 and its orthologues were measured by qPCR in indicated Bacteroidales strains. Large intestinal content (Li-Cont) and mucus (Li-Muc) were collected 4 wk after colonization of SPF WT mice previously treated with antibiotics (SPF-Abx mice; n = 3–11 mice). The relative values of Li-Muc in comparison with Li-Cont are plotted in the graph. Each plot in B and E and each column in C represents the result obtained from independent mice. Statistical analyses were performed with Student’s t test in B and E. *P < 0.05 and ***P < 0.001. Error bars represent SEM.

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