Macrophage accumulation and activation is reduced in vismodegib-treated mice on the FFC diet.

<p>(A) Total RNA was extracted from liver tissue obtained from mice treated as described in Fig. 1 and expression profile of several macrophage markers was evaluated by real-time PCR. (B) Another marker of macrophages, Mac-2, was examined by immunohistochemistry on paraffin-embedded liver tissue and representative microphotographs taken with a 20× objective are shown. Macrophage accumulation was assessed by morphometric analysis of Mac-2 positive area in ten random fields per liver tissue section as illustrated in the right panel. (C) Gene expression of cytokines related to macrophage activation, IL-1β, IL-6 and MCP-1, was analyzed by real-time PCR in liver tissue obtained from each experimental group. (D) Liver macrophages were isolated from mice on chow and the FFC diet treated with vehicle or vismodegib. Total RNA was extracted and gene expression of hedgehog signaling target genes were assessed by real-time PCR. Bar columns represent mean ± S.E.M. *** <i>P</i><0.001, ** <i>P</i><0.05, * <i>P</i><0.01.