A939572

HSCs were also isolated from mice having the third exon of the Scd2 gene flanked by loxP sites (Scd2f/f). Scd2 ablation was achieved in culture by infecting the cells with adenovirus expressing Cre recombinase. Scd2f/f mice were crossed with α1(I)collagen promoter-Cre (CC) mice to generate the conditional KO mice (Scd2f/f;CC). Scd2f/f and Scd2f/f;CC mice were given phenobarbital (0.025%) in drinking water and injected with CCl₄ (s.c., 1–2 μl/g) in mineral oil (1:1 dilution) or mineral oil only twice weekly for 4 wk. The mice were also subjected to aseptic BDL and liver fibrosis assessed on day 13. Pharmacologic inhibition of SCD with A939572 (BioVision, Inc.) was tested in CCl₄ and BDL models. Livers were collected for morphometric analysis of Sirius red, reticulin staining, and immunoblot analysis of ACTA2. Blood was collected to measure plasma ALT activity as per assay manufacturer’s (Sigma) protocol. Hydroxyproline content of livers was determined by hydrolysis of liver proteins with HCl and ELISA assay (Cell Biolabs, Inc.). HSCs were isolated from 3 pairs of Scd2f/f;CC vs. Scd2f/f mice subjected to BDL for Scd2 and Scd1 mRNA qPCR, revealing selective 89% reduction in Scd2 in Scd2f/f;CC.