Figure 7.

Exposure of Aged Mice to a Young Microbiota or Flagellin Restores Small Intestinal Crypts

(A) Olfm4 mRNA expression was compared by RT-qPCR 24 h after flagellin stimulation of in vitro enteroids prepared from small intestinal crypts. Horizontal line, median. n = 3/group. Statistical difference determined by t test.

(B) IHC detection of OLFM4 (red) in small intestinal crypts of young, aged, and aged mice given young bedding. Nuclei detected with DAPI (blue). Scale bar, 50 μm.

(C and D) Quantitation of the number of OLFM4+ cells in small intestinal crypts (C) and the crypt base (D) from mice in each group in (B) Each point is the mean of an individual mouse. n = 3–4/group. Horizontal line, median. Statistical differences determined by one-way ANOVA.

(E) IHC detection of OLFM4 in small intestinal crypts from PBS- and flagellin-treated aged mice. Nuclei detected with DAPI (blue). Scale bar, 50 μm.

(F and G) Quantitation of the number of OLFM4+ cells in small intestinal crypts (F) and the crypt base (G) from mice in each group in (E). Each point is the mean of an individual mouse. n = 3–4/group. Horizontal line, median. Statistical differences determined by t test.

(H) IHC detection of pS6 (red) in small intestinal crypts from young, PBS-treated aged, and flagellin-treated aged mice. Nuclei detected with DAPI (blue). Scale bar: upper panels, 50 μm; lower panels, 20 μm.

(I) Quantitation of the % area pS6+ immunostaining in small intestinal crypts of mice in each group in (H). Each point is the mean of an individual mouse. n = 3–4/group. Horizontal line, median. Statistical differences determined by t test.