Panel A shows immunofluorescence images of liver sections stained for DLK1, Albumin, and nuclei during different developmental stages. Panel B shows immunofluorescence images of liver sections stained for EpCAM, Albumin, and nuclei across developmental stages. Panel C shows immunofluorescence images of liver tissue stained for E-cadherin, N-cadherin, HNF4α, and nuclei during liver development. Panel D shows fluorescence images of HNF4 alpha nuclear localization in hepatoblasts and hepatocytes at different developmental stages. Panel E shows immunofluorescence images of adult liver tissue stained for E-cadherin, N-cadherin, ZO-1, HNF4 alpha, and nuclei around portal and central veins. Panel F shows immunoblot images, protein sequence comparisons, and a bar graph analyzing relative pan-cadherin antibody affinities for E-cadherin and N-cadherin. Panel G shows immunoblot images and bar graphs comparing raw and normalized E-cadherin and N-cadherin expression levels in Can 10 cells. Panel H shows immunoblot images and a stacked bar graph analyzing p120-catenin isoform expression after siRNA treatment in cultured cells.
Expression profiles of DLK1, albumin, EpCAM, HNF4α, E-cadherin, and N-cadherin in hepatoblasts and hepatocytes during mouse liver development, and expression of E-cadherin, N-cadherin, and p120-catenin in Can 10 cells, related to Figs. 1, 2, and 9. (A and B) Expression patterns of DLK1 and albumin (A) and EpCAM and albumin (B) during mouse liver development. Liver sections from different developmental stages were immunostained with antibodies against DLK1 and albumin (A) or EpCAM and albumin (B), along with DAPI staining. (C and D) Expression of HNF4α in mouse hepatoblasts and hepatocytes during liver development. Liver sections from different developmental stages were immunostained using a combination of antibodies against E-cadherin, N-cadherin, and HNF4α, along with DAPI staining (C) or using an anti-HNF4α antibody and DAPI (D). Note that HNF4α localizes within nuclei throughout liver development. (E) Expression patterns of E- and N-cadherin in adult hepatocytes. Liver sections from P60 were immunostained with antibodies against E-cadherin, N-cadherin, ZO-1, and HNF4α, along with DAPI staining. Note the different distributions of E-cadherin and N-cadherin along PVs and CVs at P60. (F) Relative affinities of the anti-pan-cadherin antibody for E- vs. N-cadherin. Left: lysates from cells expressing GFP-tagged E- or N-cadherin were immunoprecipitated using anti-GFP antibody and analyzed by immunoblotting using anti-GFP and anti-pan-cadherin antibodies. Right: top—antigen information for the anti-pan-cadherin antibody (71-7100) and corresponding regions in E-cadherin/CDH1 and N-cadherin/CDH2. Bottom—pulled-down proteins were normalized to GFP intensity, and the ratio of the pan-cadherin signal to GFP intensity was calculated. The E-cadherin ratio is set to 1.0. Data represent means ± SD from three independent experiments. (G) Normalization of E- and N-cadherin expression in Can 10 cells using the anti-pan-cadherin antibody. Top: raw expression data; the intensity of E-cadherin is set to 1.0. Bottom: normalized values calculated using the relative antibody affinities determined in F. Data represent means ± SD from three independent experiments. (H) Immunoblot analysis of p120-catenin isoform expression in Can 10 cells. Top: lysates from cells transfected with control or p120-1 siRNA were analyzed using antibodies against p120-1 and α-tubulin. MWs of marker proteins are indicated in kDa. Bottom: quantification of p120-1 vs. other p120 isoforms in Fao and Can 10 cells. Data represent means ± SD from three independent experiments. Scale bars, 10 µm (C and the middle and bottom rows in E), 20 µm (D), 100 µm (A, B, and the top row in E). MWs, molecular weights; PVs, portal veins; CVs, central veins. Source data are available for this figure: SourceData FS1.