Extended Data Fig. 8: OGDH downregulation increases secretory-lineage specification in vivo.
From: Metabolic adaptations direct cell fate during tissue regeneration
a, Intestine-specific expression of GFP in TRE-shRenVillin-rtTA3 mice. Representative images of GFP fluorescence of various organs from TRE-shRenCag-rtTA3 mice and TRE-shRenVillin-rtTA3 mice fed a doxycycline diet for 6 days. b, H&E, ABP staining and immunofluorescence of GFP, OGDH, Ki67, lysozyme (Lyz), and 5hmC in intestinal tissues after 5 days of doxycycline treatment in TRE-shOgdhVillin-rtTA3 and TRE-shRenillaVillin-rtTA3 mice. Dashed lines outline crypt structure. c, Design of RNA-seq experiments in isolated intestinal crypts. Created in BioRender. Chaves-Perez, A. (2025) https://BioRender.com/d8uy3en. d, Top upregulated and downregulated pathways in TRE-shOgdhCag-rtTA3 versus TRE-shRenCag-rtTA3 mice from GO analysis. e, Differential transcription in crypts from TRE-shOgdhCag-rtTA3 and DM-αKG-treated mice compared to TRE-shRenCag-rtTA3 and vehicle-treated mice respectively. The heat map shows genes associated with the secretory lineage, absorptive lineage (zone 1 and zone 2), Notch signalling pathway, and cell proliferation. Each column represents a single mouse. f, Top upregulated pathways in intestinal crypts of DM-αKG-treated versus vehicle-treated mice, derived from GO analysis. g, Heat map depicting expression of genes in the WNT pathway in crypts from TRE-shOgdhCag-rtTA3, TRE-shRenCag-rtTA3, vehicle-treated and DM-αKG-treated mice after 3 days of treatment. Each lane represents one mouse. h, Heat map depicting expression of transcription factors involved in lineage specification in crypts from TRE-shOgdhCag-rtTA3, TRE-shRenCag-rtTA3, vehicle-treated, and DM-αKG-treated mice at day 3 of treatment. i, Plot showing predicted transcription factors commonly upregulated (red dots) or downregulated (blue dots) in mice treated with TRE-shOgdhCag-rtTA3 versus DM-αKG. j, Schematic depicting the dual role of OGDH in differentiation in intestinal homeostasis. Created in BioRender. Chaves-Perez, A. (2025) https://BioRender.com/7osfp6n. This figure is adapted from our published patent (WO2024229094A1)50.
