Journal: Nature Communications
Article Title: Hepatic stellate cell stearoyl co-A desaturase activates leukotriene B4 receptor 2 - β-catenin cascade to promote liver tumorigenesis
doi: 10.1038/s41467-023-38406-8
Figure Lengend Snippet: a A heatmap of DEGs in tumor-adjacent liver (TAL) of Scd2 f/f ;Col1a1Cre (CC) vs. Scd2 f/f mice subjected to the DEN + WAD regimen ( n = 3 mice per group). b IB analysis of TAL proteins from Scd2 f/f ;CC vs. Scd2 f/f mice as compared to Scd2 f/f control mice. ( n = 3 mice per group). c IF microscopy of nYAP1 + HNF4A + hepatocytes, nYAP1 + ACTA2 + aHSC, nYAP1 + SOX9 + ductular cells in Scd2 f/f ;CC vs. Scd2 f/f mouse TAL. Scale bar = 50 μm. White arrows point the cells with HNF4A + , ACTA2 + , or SOX9 + staining while yellow arrows point the cells with the dual staining with nYAP1. d Morphometric analysis of nYAP1 + hepatocytes, aHSC, and ductular cells. * p < 0.05 and ** p < 0.01 vs. Scd2 f/f mouse TAL by two-sided t-test. Data presented as means ± SEM ( n = 3 different sections). Exact p values are shown in the . e Co-IF microscopy of nYAP1 + HNF4α + liver tumor cells (HCC). Scale bar = 100 μm. The border between HCC and non-tumorous (NT) areas is indicated by a broken line. Images shown are representative of four pairs of samples analyzed. f Imaging morphometric data for the percentage of nYAP1 + HNF4α + liver tumor cells by 3-dimensional confocal microscopy analysis. * p < 0.05 vs. Scd2 f/f by two-sided t-test. Data shown are means ± SEM ( n = 4 pairs of samples). g qPCR data for Scd2 f/f ;CC vs. Scd2 f/f mouse TAL ( n = 6 each) compared to Scd2 f/f control normal liver ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.005 vs. Scd2 f/f control; # p < 0.05 and ## p < 0.01 vs. Scd2 f/f TAL by two-sided t-test. h Lipidomic analysis for PUFA metabolites in Scd2 f/f ;CC vs. Scd2 f/f mouse TAL. Data presented as means ± SEM ( n = 3 mouse samples per group). P values determined by two-sided t-test. Red arrows depicting reductions in four specific metabolites in Scd2 f/f ;CC TAL (Raw data are provided in ). i IB analysis of LTB4R2 for Scd2 f/f ;CC ( n = 6 mice) vs. Scd2 f/f ( n = 5 mice) TAL proteins compared to Scd2 f/f control mice ( n = 3 mice). For all relevant figures, source data and exact p values are provided in the file.
Article Snippet: After blocking the endogenous peroxidase activity and non-specific protein binding sites, the sections were incubated with primary antibodies for rabbit-YAP (1:500) from Abcam, and YAP expressing cells were co-stained with mouse-α-SMA (1:500) from Sigma-Aldrich, goat-HNF4α (1:500) from Santa Cruz Biotechnology, mouse-SOX9 (1:1000) from Merck, followed by incubation with the fluorescent secondary antibody (Supplementary Table ).
Techniques: Control, Microscopy, Staining, Imaging, Confocal Microscopy
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