Mice with HSC selective deletion of synectin (Col^cre/Synectin^fl/fl) and their control littermates (Synectin^fl/fl) were treated with either olive oil (vehicle) or CCl₄ via i.p. injections twice a week for 6 weeks. The livers were then harvested and prepared for analysis through isolation of mRNA or protein, or fixation of liver tissue for immunostaining and Sirius red analysis. (A) qPCR for collagen-1 mRNA levels showed a significant reduction in Col^cre/Synectin^fl/fl mice liver after CCl₄ injection, n = 4–7 per group. (B) Collagen content was reduced in Col^cre/Synectin^fl/fl mice after CCl₄ injection, as demonstrated by assessing hepatic hydroxyproline assay, n = 4–7 per group. (C) Liver sections (5 μm) were stained with Sirius red to represent the fibrotic strands correlating with the degree of fibrosis. Additional liver sections were stained with antibodies against the fibrotic proteins collagen-1, PDGFR-α, PDGFR-β, and α-SMA (green) in conjunction with nuclear costaining with DAPI (blue), representative images shown. Staining revealed decreased expression of collagen-1, PDGFR-α, PDGFR-β, and α-SMA in Col^cre/Synectin^fl/fl mice after CCl₄ injection compared with control littermates. Quantitation was performed using ImageJ, with fold change displayed on the micrographs and graphs located in Supplemental Figure 2, A–E; n = 3–5. Scale bars: 200 μm. (D) Lysates from whole mouse liver were used to assess protein levels of PDGFR-α, PDGFR-β, and synectin in Col^cre/Synectin^fl/fl mice after CCl₄ injection. Samples were run on the same gel but were noncontiguous. n = 3–4. Quantification was performed using ImageJ, with the densitometric values displayed below the blots and graphs located in Supplemental Figure 2F. All data are displayed as mean ± SEM (*P < 0.05). Each dot in the scatter plot indicates an individual animal in each of the panels. One-way ANOVA with Bonferroni’s multiple comparison tests were used to analyze groups for statistical significance (*P < 0.05, **P < 0.001, ***P < 0.0001).