Fig. 2: CREPT deletion reduces the Lgr5+ cell number.
From: CREPT is required for murine stem cell maintenance during intestinal regeneration
a A quantitative RT–PCR analysis of the expressions of CBC and reserve stem cell markers in WT and Vil-CREPTKO crypt cells. b Representative images of Lgr5-GFP+ cells in WT and Vil-CREPTKO intestines. c Quantification of Lgr5-GFP + cells in small intestines of WT and Vil-CREPTKO mice based on FACS analyses. n = 3 independent experiments. p = 0.0042. d Representative images of Lgr5 and CREPT staining in intestine crypts. e A FACS analysis of Lgr5 and CREPT stained cell populations. The population of crypts cells, which had the highest level of Lgr5, also expressed the largest amount of CREPT protein. f A quantitative RT-PCR analysis of Lgr5 and CREPT in sorted Lgr5-GFP+ and Lgr5-GFP− cells. n = 3 independent experiments. p < 0.0001 (Lgr5 in Lgr5− vs. Lgr5+ cells). p = 0.0002 (CREPT in Lgr5− vs. Lgr5+ cells). g A schematic diagram showing the TAM treatment in Lgr5-GFP; CREPTfl/fl mice. h Representative images of Lgr5-GFP staining in intestines at indicated time post TAM treatment. i Percentage of Lgr5-GFPhigh cells in crypts at indicated time post TAM treatment by FACS analyses. p = 0.0393 (day 1 vs. day 7). p = 0.0011 (day 1 vs. day 30). j Representative images of WT and CREPT deleted organoid formation derived from Lgr5-GFP and Lgr5-GFP; CREPTfl/fl mice. k The numbers of WT and Lgr5-CREPTKO organoids showing <100 μm and ≥100 μm diameters. Images are representative of at least three independent experiments. Statistics data represent mean ± SEM. All p values were generated by 2-tailed Student’s t-test. Scale bars: 100 μm (b, h, j), 10 μm (d).
