Figure 3 from SALL4 Is Required for YAP1-Dependent Malignant and Regenerative Hepatocyte-to-Cholangiocyte Reprogramming

<p><i>Sall4</i> overexpression disrupts <i>YAP1</i>-driven HC-to-CCA transformation while enhancing the clonal expansion of fatty LPC-like cells. <b>A,</b> Schematic representation of the experimental design. FVB mice underwent HDTVI with transposon vectors expressing HA-tagged <i>myrAkt</i> and <i>Yap1</i><sup><i>S127A</i></sup> (<i>AY</i>) combined with GFP expression (<i>AY-GFP</i>) or V5-tagged <i>Sall4</i> (<i>AY-Sall4</i>) plasmids and were analyzed at indicated time points (2.5 and 5 weeks after HDTVI). <b>B,</b> qRT-PCR showing <i>Sall4</i> expression in <i>AY-Sall4</i> compared with <i>AY-GFP</i>. <b>C,</b> Representative liver images from <i>AY-GFP</i> or <i>AY-Sall4</i> mice at 2.5 (top) and 5 weeks (bottom) after HDTVI. <b>D,</b> LW/BW ratio analysis comparing <i>AY-GFP</i> and <i>AY-Sall4</i> at indicated time points. <b>E</b> and <b>O,</b> IHC analysis for indicated markers (HA-tag, panCK, HNF4A, Ki-67, and V5-tag) at indicated time points. <b>F–N,</b> Quantification of indicated parameters [tumor area, panCK<sup>+</sup> area, lipid droplet area, Ki-67<sup>+</sup> cells, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), bilirubin, CD45, TUNEL, and α-SMA] comparing <i>AY-GFP</i> and <i>AY-Sall4</i>. 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> < 0.001; ****, <i>P</i> < 0.0001). Scale bars, (<b>E</b> and <b>O</b>) 1 mm (overview), 200 μm (high magnification); (<b>K</b>) 200 μm. ns, not significant.</p>