Figure 1. ISC-specific deletion of CDC42 causes crypt hyperproliferation.
Two- to 3-month-old CDC42 ISC-KO (Olfm4-CreER, CDC42flox/flox) and control (Olfm4-CreER, CDC42flox/+) mice were injected with TAM once per day for 3 days, and then were sacrificed at 12 h (day 0.5), 24 h (day 1), 72 h (day 3), or 96 h (day 4) after the third TAM injection.
(A) Small intestinal crypts were isolated, lysed, and western blotted for CDC42. Actin is the loading control. (Upper) Day 1 and day 4 whole-crypt cell lysates. (Lower) Day 1 GFP sorted crypt cells.
(B and C) Representative images of H&E staining of duodenal sections; one crypt is circled in each image based on morphology.
(D–M) Representative images of immunofluorescence staining of duodenal sections.
(D and E) Anti-pH3 and anti-E-cadherin; one crypt is circled in each image.
(F–K) Lineage tracing tdTomato, DAPI, E-cadherin, as well as lysozyme, Ki67, and Olfm4; one crypt is circled in each day 3 image.
(L and M) anti-αE-catenin; enlargements of villi (Ľ and M’) and crypt (Ľ’ and M”).
Data are representative of at least three independent experiments. Scale bars, 50 μm. n = 4 for each genotype.