Fig. 2: MTGR1 loss increases ISC number and deregulates intestinal stem cell programs.

A Immunofluorescent staining for Ki67 (red), E-cadherin (green), and nuclei (DAPI, blue) in the small intestine of 8–12-week-old WT and Mtgr1^−/− mice. n = 4 mice per genotype, >20 high-powered fields (HPFs) per mouse. B Lgr5 mRNA expression was visualized in the WT and Mtgr1^−/− small intestine by RNAscope. n = 4 WT and 3 Mtgr1^−/− mice, 12 HPFs per mouse. C Mtgr1^−/− mice were intercrossed with the Lgr5-cre-EGFP reporter strain and isolated crypt cells were stratified by Lgr5-EGFP expression through FACS. n = 4 WT and 3 Mtgr1^−/− mice. D Lgr5-EGFP assessed by immunofluorescence. Quantification shows the number of GFP positive cells in each reporter positive crypt, per mouse. n = 9 mice per genotype. E q-RT-PCR of stem cell markers Lgr5, Myc, Ki67, Ascl2, and Olfm4 in intestinal crypt isolates from Mtgr1^−/− and WT mice (n = 3–6 mice per genotype). Results were normalized to Gapdh and represented as fold change over WT expression. F UMAPs depicting cell types as determined from scRNA-sequencing results from WT and Mtgr1^−/− intestinal cells and (G) numerical representation. n = 3 WT and 2 Mtgr1^−/− duodenal samples. ABS absorptive, EE enteroendocrine, GOB goblet, PAN Paneth, RevSTM revival stem, STM stem, TAC transit amplifying cell, TUF tuft. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, Mann–Whitney test (A, D, select E), Student’s t test (B, select E), or two-way ANOVA with Sidak’s multiple comparison test (C), Scale bars = 100 µm.