Fig. 4. Muscular Mmp17 regulates crypt formation.

a, b Lgr5/Olfm4 RNAscope representative images of WT vs KO intestines and their quantification. Scale bar 100 µm; 25 µm in inset. n = 6 mice per genotype in Colon Lgr5 and WT Olfm4 and 7 mice per genotype in the rest. c Representative bright-field images of colonoids from WT vs KO crypts 48 h after crypt isolation (left). Scale 650 µm; 50 µm in cropped image. Graph shows relative colonoids number as percentage normalized to WT. n = 5 experiments performed. One representative experiment is showed in which each dot represents a well.,. d Graphs represent the percentage of colonoids after the first split (left) and the percentage of colonoids derived from colon crypts after 72 h in response to the enriched medium WNR (right). n = pooled 4 wells per condition in ENR WNT experiments and 6 wells per condition in WNR experiment performed in two independent experiments. e Schematic of crypt formation assay on iECM; SI organoids were seeded on the iECM, counted on day 1 for attachment and survival and left to engraft into the scaffold until Day7. de novo crypts per seeded organoid (as assessed at day 1) were counted at day 7 post plating. The number of de novo crypts was counted by brightfield imaging as shown in (f). f Representative brightfield (top) and fluorescent (bottom, E-Cadherin (red), DAPI (blue), images showing epithelial crypt formation in WT and KO iECM. Arrows demonstrate the de novo formed crypts. Scale bar 50 µm (brightfield) and 20 µm (fluorescent). n = 2 (fluorescent) to 5 (brightfield) mice per genotype. g Quantification of number of crypts in WT and KO iECM. n = 4 WT and 5 KO mice pooled from two independent experiments (based on brightfield imaging). Numerical data are means ± SD and were tested by Mann–Whitney test (two-sided, with the following p values; p = 0.0411 (colon) and p = 0.0140 (SI) in (a); p = 0.0082 in (b) and p = 0.0378 in (c)) and Student’s unpaired t-test (two-tailed, p < 0.0001) in (g). Source data are provided as a Source Data file.