Figure 5. Maternal diet impacts microbiota localization and pro-inflammatory potential in offspring.
(A-B) Microbiota-mucus-epithelial localization in colon of 12-week-old offspring via FISH. Representative confocal images (A) and average bacterial-epithelial distances per HPF (B).
(C&D) Bacterial IgA coating. Gating scheme in (C), using Rag−/− feces as neg control, was used to discern the percentage of fecal bacteria coated with IgA (D).
(E-M) Flow cytometric analysis of intestinal immune cells in 12-week-old offspring (N=5) as gated in Figure S1, Neutrophils (Neu), monocytes, M1/M2 macrophage, Dendritic cells (DC), B and T lymphocytes were quantitated.
(N-Q) Functional T-cell subset analysis (N=3-5). Small intestinal lymphocytes and splenocytes were stimulated with PMA ex vivo and analyzed by FACS gating in (N). Percentage of regulatory T cells (O), IFN-γ+ (P), IL-17+ T cells(Q) were calculated. One-way ANOVA: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. See Figure S3&S4