Fig. 2: Altered lineage relations in the crypt translate into the mature villus compartment.

a, Foxa2^{nEGFP-CreERT2/+};Gt(Rosa26)^mTmG/+ lineage-tracing model. mT Foxa2-negative cells (red) convert into mG Foxa2-lineage-positive cells (green) upon Foxa2-promoter-driven Cre expression via an intermediate (mTmG⁺, yellow) state. pCA, chicken β-actin core promoter with a CMV enhancer. b, Experimental scheme of short-term lineage tracing of Foxa2-positive cells using the Foxa2^{nEGFP-CreERT2/+};Gt(Rosa26)^mTmG/+ mouse model. c,d, Representative laser scanning microscopy (LSM) images of Cre-driven recombination in the duodenum of CD- and HFHSD-fed Foxa2^{nEGFP-CreERT2/+};Gt(Rosa26)^mTmG/+ (c) and analysis of lineage-positive cells (d) 70 h after tamoxifen induction. Single converted mG⁺ cells and lineage ribbons (green) containing Muc2⁺ goblet cells (red), ChgA⁺ EECs (white) and villin⁺ enterocytes (white) are observed in crypts and villi. Scale bar, 100 µm. For Foxa2-lineage analysis, only confluent mG⁺ cell patches located in the villi were considered (n = 3 mice per group). Data are mean ± s.e.m. Statistical significance was determined by two-tailed Student’s t-test. DAPI, 4,6-diamidino-2-phenylindole. e–j, Abundances of mature intestinal cell types are altered in HFHSD-fed FVF mice. e,g, Representative LSM images (e) of goblet cells (Muc2⁺) and quantification thereof (f,g). Scale bar, 75 µm, n = 3 mice per group. h–j, Representative LSM images (h) of ChgA⁺ EECs and quantification thereof (i,j). Scale bar, 75 µm, n = 4 mice per group. Data are mean ± s.e.m. of biologically independent samples. Statistical significance was determined by two-tailed Student’s t-test.

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