Fig. 3: Heated-inactivated B. adolescentis supplementation improved colonoids performance in vivo.

a, b The expression of the Lgr5 gene was detected in mice colon tissue from different age groups by immunofluorescence and western blot, n = 3 animals per age group. Scale bar, 50 μm. c Representative images of colonoids derived from 3-month-old mice and 12-month-old mice with n = 3 independent experiments with similar results. Arrows indicate crypt domains. Scale bar, 100 μm. d The schematic diagram of the experimental procedure in the young (3-month-old mice) and old (12-month-old mice) groups. e Representative images are shown from one out of 4 independent experiments with similar results in the organoid-forming capacity of crypts from Young, Old+PBS and Old+B.a mice group. Scale bar, 100 μm. f Relative size of colonoids quantified on days 3, 5 and 7 and represented relative to the Young group. n = 4 independent experiments per group) on days 3, 5 and 7. Data were represented as mean ± SEM. Comparisons were performed by Two-way ANOVA analysis followed by Tukey’s multiple comparisons test. g Relative mRNA levels of the Lgr5 gene in Young, Old+PBS and Old+B.a mice group, n = 3 individual experiments. Data were represented as mean ± SEM. Comparisons were performed by unpaired, two-tailed t test. *p < 0.05, **p < 0.01, ***p < 0.001. Source data and exact p value are provided as a Source data file.