Induction of senGC maturation coincides with colitis protection. (A) Timing of colitogenic DSS challenge in relation to microbiota-dependent induction of senGC maturation in adult GF mice. (B) Tracking of day zero normalized mass changes in H2O- and DSS-treated ConvD mice from senGC− and senGC+ groups. (C) Photographs of colonic tissue dissected from different treatment groups at the termination of the DSS challenge experiment. (D) Quantification of total colon length in different treatment groups based on images shown in C. All data normalized to average colon length of the H2O control animals for each group. (E) Quantification of CLN mass in different treatment groups. All data normalized to average CLN mass of the H2O control animals for each group. (F) Quantification of spleen mass in different treatment groups. All data normalized to the average spleen mass of the H2O control animals for each group. (G) Determination of mucosal bacterial load in different treatment groups by 16S qPCR of colonic tissue DNA. (H) Determination of bacterial translocation from the colon in different treatment groups by 16S qPCR of CLN DNA. (I) Combined histology scores from different treatment groups determined by histopathological scoring of fixed colonic Swiss roll tissue sections. (J) Individual scores for different histopathology metrics from different treatment groups. (K) Micrographs of AB-PAS–stained fixed colonic Swiss roll tissue sections used for histology scoring. Areas of ulceration (asterisk) and GC depletion (yellow arrowhead) in senGC− DSS-treated tissue are indicated. (L) Ulcer coverage in DSS-treated mice as a fraction of total mucosal surface area. Data represent n = 4–7 animals per group, as indicated. All data are pooled from two independent experiments. All error-bar graphs show median and interquartile range. Statistical comparisons between groups by Kruskal–Wallis and uncorrected Dunn’s (B–J) or Mann–Whitney test (L); P < 0.05 (*), <0.01 (**), <0.001 (***), <0.0001 (****). Image scale bars are 2 cm (C) or 100 µm (K).
Figure 6.

Induction of senGC maturation coincides with colitis protection. (A) Timing of colitogenic DSS challenge in relation to microbiota-dependent induction of senGC maturation in adult GF mice. (B) Tracking of day zero normalized mass changes in H2O- and DSS-treated ConvD mice from senGC− and senGC+ groups. (C) Photographs of colonic tissue dissected from different treatment groups at the termination of the DSS challenge experiment. (D) Quantification of total colon length in different treatment groups based on images shown in C. All data normalized to average colon length of the H2O control animals for each group. (E) Quantification of CLN mass in different treatment groups. All data normalized to average CLN mass of the H2O control animals for each group. (F) Quantification of spleen mass in different treatment groups. All data normalized to the average spleen mass of the H2O control animals for each group. (G) Determination of mucosal bacterial load in different treatment groups by 16S qPCR of colonic tissue DNA. (H) Determination of bacterial translocation from the colon in different treatment groups by 16S qPCR of CLN DNA. (I) Combined histology scores from different treatment groups determined by histopathological scoring of fixed colonic Swiss roll tissue sections. (J) Individual scores for different histopathology metrics from different treatment groups. (K) Micrographs of AB-PAS–stained fixed colonic Swiss roll tissue sections used for histology scoring. Areas of ulceration (asterisk) and GC depletion (yellow arrowhead) in senGC− DSS-treated tissue are indicated. (L) Ulcer coverage in DSS-treated mice as a fraction of total mucosal surface area. Data represent n = 4–7 animals per group, as indicated. All data are pooled from two independent experiments. All error-bar graphs show median and interquartile range. Statistical comparisons between groups by Kruskal–Wallis and uncorrected Dunn’s (B–J) or Mann–Whitney test (L); P < 0.05 (*), <0.01 (**), <0.001 (***), <0.0001 (****). Image scale bars are 2 cm (C) or 100 µm (K).

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