Expression of Cyp2e1 and Alb mRNA in livers after CCl4 challenge by in situ hybridization. Liver sections were subjected to in situ hybridization with specific probes for Cyp2e1 and Alb genes. (A) Double in situ hybridization showed staining of Cyp2e1-expressing cells as a deep blue color stained with NBT/BCIP-substrate bordering the injured areas (indicated by the arrows), mainly in the pericentral vein area. Alb expression, shown in deep brown stained with INT/BCIP-substrate, was observed throughout the liver, except in the pericentral vein area, at 3 days and 36 days after CCl4 challenges in both WT and PF4CreTgfb1f/f mice. (B) Quantification of Cyp2e1 and Alb expression showed cells expressing the Cyp2e1 gene in the injured areas around the pericentral veins in both WT and PF4CreTgfb1f/f mice at 3 days after CCl4 challenge. (C) Gpx4 expression in the liver was assessed by in situ hybridization using NBT/BCIP-substrate and showed similar expression levels in both WT and PF4CreTgfb1f/f mice at 6 hours after CCl4 challenge. (D) Quantification of Gpx4 6 hours after CCl4 challenge in WT and PF4CreTgfb1f/fmice.
