Figure 5 from SALL4 Is Required for YAP1-Dependent Malignant and Regenerative Hepatocyte-to-Cholangiocyte Reprogramming

<p>SALL4 regulates regenerative HC-to-CC conversion in the DDC-fed cholestasis model. <b>A,</b> Schematic experimental design. <i>Sall4</i><sup><i>fl/fl</i></sup> mice were injected with either <i>AAV8-TBG-GFP</i> (<i>Sall4 WT</i><sup><i>hep-GFP</i></sup>) or <i>AAV8-TBG-Cre</i> (<i>Sall4 KO</i><sup><i>hepΔGFP</i></sup>) 2 weeks prior administration of a DDC-containing diet for 4 weeks. Liver tissues were analyzed at the end of the treatment period. <b>B,</b> LW/BW ratio comparison and serum biochemistry analysis comparing alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bilirubin levels between <i>Sall4 WT</i><sup><i>hep-GFP</i></sup> and <i>Sall4 KO</i><sup><i>hepΔGFP</i></sup> groups (<i>n</i> = 4–6 mice per group). <b>C,</b> Representative images of CD45 IHC and Sirius Red staining in livers sections from both groups. <b>D</b> and <b>E,</b> Quantification of (<b>D</b>) CD45<sup>+</sup> inflammatory cells per 1,000 μm<sup>2</sup> and (<b>E</b>) collagen deposition (%) by Sirius Red staining. <b>F,</b> Representative liver sections stained with hematoxylin and eosin (H&E), SOX9 (CC lineage marker), and Ki-67 (proliferation marker). <b>G</b> and <b>H,</b> Quantification of (G) SOX9<sup>+</sup> cells (%) and (<b>H</b>) Ki-67<sup>+</sup> cells in the periportal area (%). <b>I,</b> IF staining for SOX9, HNF4A, and GFP. Yellow arrowheads indicate intermediate cells expressing both CC (SOX9) and HC (HNF4A) markers. <b>J,</b> Quantification of SOX9<sup>+</sup>HNF4A<sup>+</sup> double-positive intermediate cells among total HNF4A<sup>+</sup> cells (%). <b>K,</b> IF staining for CK19 and GFP to visualize HC-derived CC. <b>L,</b> Quantification of CK19<sup>+</sup> CCs per microscopic field. <b>M,</b> Quantification of CK19<sup>+</sup>GFP<sup>+</sup> double-positive cells among total CK19<sup>+</sup> cells (%). Data are presented as the mean ± SD; Student <i>t</i> test (*, <i>P</i> < 0.05; **, <i>P</i> < 0.01; ****, <i>P</i> < 0.0001; ns, not significant). Scale bars, (<b>C–F</b>) 100 μm; (<b>K–M</b>) 50 μm.</p>