Fig. 6. icGC alterations in Spdef^−/− colon.

(A) Fold change (FC) expression of single GC cluster (color bars) marker genes in Spdef^−/− versus WT with adjusted p-values. (B) FC expression among the previously defined single cell clusters (left) of the most up- and down-regulated genes in Spdef^−/− compared to WT GCs obtained by bulk RNA seq with adjusted p-values (right). (C) UEA1, cell membrane and DNA staining of whole-mounted colonic tissue from 7 and 13 week-old WT and Spdef^−/− mice. Magnified areas (white boxes), apical surface of icGCs (arrowheads), scale bars = 20 μm. (D) Quantification of the number of icGCs per mm² in whole-mounted colons from WT and Spdef^−/− mice. Mean ± SD, analyzed by 2-way ANOVA with Sidak’s multiple comparison test; n= 4 - 8. (E) TEM micrographs of icGCs in WT and Spdef^−/− colon with magnifications. Scale bars = 2 μm; n = 4. (F) SEM scans of the icGCs and the epithelial surface in WT and Spdef^−/− distal colon. Identified GC openings (red arrowheads), scale bars = 1 μm (upper) 20 μm (lower), n = 4. (G) In vivo labeling for 2 and 4 h by GalNAz detected by Alkyne-TAMRA on colonic sections of 13-week old WT and Spdef^−/− mice. Scale bar = 20 μm; n = 4.