Characterization of Dpt ^creER ;R26 ^tdTomato and Acta2-creER;R26 ^tdTomato mice. (A) Schematic of the design of Dpt^creER;R26^tdTomato lineage-tracing mice. (B) Quantification of the percentage of CD34⁺ fibroblasts that were Tomato⁺ on the liver sections from Dpt^creER;R26^tdTomato mice. (C–E) On-slide staining of non-parenchymal liver cells from Dpt^creER;R26^tdTomato mice with anti-Col1 (C), anti-Myh11 (D), and anti-Reln (E) antibodies, respectively. Cells were isolated by gradient centrifugation and were mounted on poly-D-lysine–coated slides. Scale bar = 25 µm. (F and G) Confocal images showing that Tomato was not expressed in Hnf4a⁺ hepatocytes (F) or F4/80⁺ Kupffer cells (G) in the livers from Dpt^creER;R26^tdTomato mice. Scale bar = 50 µm (n = 5 mice from three independent experiments). (H) Statistical analysis of the percentage of CAFs derived from Cd34-CreER⁺ and Dpt-CreER⁺ cells in ICCs. The statistical significance of differences was assessed by two-tailed Student’s t tests. ***P < 0.001. (I) Confocal images of liver sections (15 µm) showing the protein expression patterns of aSma. Blood vessels and bile ducts were labeled by anti-CD31 and anti-Epcam antibodies, respectively. Scale bar = 200 µm (n = 3 mice from three independent experiments). (J and K) High-magnification confocal images showing the distributions of Tomato⁺ cells and Col1⁺ cells in the livers from Acta2;R26^tdTomato mice. J represents a magnified view of Fig. 5 E. Scale bar = 50 µm (n = 3 mice from three independent experiments). (L) Confocal images showing that Tomato was not expressed in Hnf4a⁺ hepatocytes in the liver from Acta2;R26^tdTomato mice. Scale bar = 50 µm (n = 3 mice from three independent experiments).