Fig. 3.

Maternal restraint stress alters IgA binding to offspring fecal microbiota in a sex-specific manner. (a) Stool from 2-week-old pups of the indicated sex born from control or stressed dams was stained with anti-IgA antibodies and analyzed by flow cytometry. Stool samples from Rag1−/− and germ-free mice were included to assess the degree of anti-IgA antibody nonspecific binding and bacterial contamination, respectively. Only populations positive for SYTO BC, a nucleic acid stain, were included in the analysis. Cumulative data is shown at top right. **p < 0.01, Student’s t test. n = 8 pups per group. Data are pooled from two independent experiments. Horizontal bars show mean + SD. (b) Mean fluorescent intensity (MFI) of IgA⁺ bacteria. **p < 0.01, Student’s t test. n = 8 pups per group. Data are pooled from two independent experiments. Horizontal bars show mean + SD. (c) Linear regression modeling of fecal supernatant IgA levels correlating with percentage of IgA⁺ fecal bacteria. Blue and red dots denote pups from non-stressed and stressed dams, respectively. r² and P values were calculated with Pearson’s r correlation test. Data are pooled from two independent experiments. n = 8 pups per group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)