Fig. 6: Pericryptal loss of PRC2 alters stem cell homeostasis.

a Schematic illustrating the loss of PRC2 in pericryptal cells comprising the intestinal stem cell niche. b GFP expression in Pdgfrβ^Cre-ERT2/+;Rosa26^mTmG mice upon induction with tamoxifen (100 mg/kg, 20 mg/mL). c–e Pdgfrβ^+/+;Eed^fl/fl (left) and Pdgfrβ^Cre-ERT2/+;Eed^fl/fl (right) (n = 3 biologically independent samples for each genotype). c Haematoxylin and Eosin staining of duodenal sections. Black dotted lines outline the crypts of each mouse. Crypt length (µm) was quantified by measuring the longest edge of each crypt using Fiji. Each point represents an individual crypt measurement (n = 4 Pdgfrβ^+/+;Eed^fl/fl mice and n = 5 Pdgfrβ^Cre-ERT2/+;Eed^fl/fl mice, ±SEM, ***P < 0.001 by Student’s t-test, two-tailed). Source data are provided as a source data file. d OLFM4 immunofluorescence identifying stem cells in the crypt base. Number of OLFM4-positive cells were quantified per crypt, with each dot representing a single measurement (duodenal tissues, n = 3 biologically independent samples for each genotype, ±SEM, ***P < 0.001 by Student’s t-test, two-tailed). Source data are provided as a source data file. White dashed lines separate the epithelium (Epi) and stromal cells (Str). All scale bars represent 50 µm. e Differential expression of all secreted ligands following Eed KO in adult and embryonic intestinal mesenchymal cells. * denotes significantly DE in between WT and Eed KO at the respective age. *p < 0.05; **p < 0.01; ***p < 0.001. f Correlation of expression of secreted ligands (log(TPM + 1)) comparing E14.5 WT Int (x-axis) versus WT Adult Int, E14.5 KO Int, and E14.5 WT St (y-axis; green, red, and blue dots, respectively). Ligands shown are as in e. Genes identified as PRC2-sensitive in E14.5 WT Int are labeled. TPM values were calculated from spike-in normalized counts and averaged between replicates. Spearman’s rho is reported for each axis.