(A) Mucus thickness in mouse ileum immediately after
explanation (time 0), after mucus removal (MR) at 20 min, and at the indicated
time points after the tissue was stimulated by basal perfusion with carbachol
(Cch) at 40 minutes. Data presented as mean ± SD (n = 5 independent
experiments). (B) Immunofluorescence staining for Muc2 in goblet
cells in the crypts of unstimulated (Unstim) and Cch-stimulated ileal explant
tissue. Scale bar, 10 μm. Image is representative of 3 independent
experiments. (C) Multi-photon live tissue imaging of small
intestinal crypts from RedMUC298tr mice) after the tissue was
stimulated with Cch at time 0. Scale bar, 10 μm. Images are
representative of n = 3 independent experiments. (D) Quantification
of theca volume of individual mCherry+ cells from the crypt in panel
C. (E) Live confocal imaging of ileal organoids generated from
RedMUC298tr mice and stimulated with acetylcholine (Ach). Scale
bar, 50 μm (main images) and 10 μm (insets). Images are
representative of n = 3 independent experiments.