Fig. 6: dediHeps give rise to ISC-like cells following forced expression of Cdx2. | Nature Communications

Fig. 6: dediHeps give rise to ISC-like cells following forced expression of Cdx2.

From: Hepatocytes differentiate into intestinal epithelial cells through a hybrid epithelial/mesenchymal cell state in culture

Fig. 6

a Representative micrographs of 3D cultures of dediHeps transduced with GFP or Cdx2 (with GFP). Initially, dediHeps transduced with GFP or Cdx2 were embedded in Matrigel and cultured with WCENR. After passage 5, dediHep-derived SOs and BOs were cultured with ENR to promote the spontaneous transition of SOs to BOs. Crypts were consistently cultured with ENR. b qPCR analyses of Cdx2 expression by dediHeps transduced with GFP or Cdx2 (with GFP) 1 day (d) and 3 d after viral infection. All data were normalized to the value for dediHeps transduced with GFP at 1 d and are depicted as fold-changes. c Co-immunofluorescence staining (GFP with Cdx2) of dediHeps transduced with Cdx2 (with GFP) 1 day after viral infection. d Volcano plots depicting differences in gene expression between hepatocytes and dediHep-derived BOs or ISC-derived BOs. Each dot represents one gene. Blue and red dots represent down- and up-regulated genes, respectively, in dediHep-derived BOs or ISC-derived BOs. Gray dots represent genes that are not significantly differentially expressed. e A Venn diagram shows direct comparisons of the global gene expression profiles between dediHep-derived BOs and ISC-derived BOs. f Co-immunofluorescence staining (E-cad with Muc2, Villin, ChgA, Lyz, or CC3 and Sox9 with EphB2) of dediHep-derived BOs and ISC-derived BOs. Insets, representative ChgA+ E-cad+ cells. g qPCR analyses of expression of Cdx2 and Vil1 and marker genes for differentiated intestinal epithelial cells (Lyz1, Chga, and Muc2) and ISCs (Lgr5 and Ascl2) in hepatocytes freshly isolated from adult mouse livers, dediHeps maintained in monolayer culture, SOs formed by dediHeps transduced with GFP, BOs formed by dediHeps transduced with Cdx2 (with GFP), and ISC-derived BOs. All data were normalized to the values for ISC-derived BOs and are depicted as fold-changes. DNA was stained with DAPI. Scale bars, 200 μm (a) and 50 μm (c, f). Data represent means ± SEM (n = 3 independent experiments). Statistical difference was determined by two-sided Student’s t test (b), quasi-likelihood F-test (d), or one-way analysis of variance followed by Dunnett’s multiple comparison test (g). Source data are provided as a Source Data file.

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