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. Author manuscript; available in PMC: 2017 Dec 13.
Published in final edited form as: Cell Metab. 2016 Oct 27;24(6):848–862. doi: 10.1016/j.cmet.2016.09.016

Figure 5. TAZ Induces Ihh, and TAZ Silencing Lowers the Expression of Pro-Fibrotic Hedgehog Pathway Genes in the Livers of FPC-Fed Mice.

Figure 5

(A) Conserved TAZ/TEAD consensus sequence in intron 1 of the mouse Ihh gene.

(B) Liver nuclear extracts from mice fed chow diet or FPC diet for 16 weeks with or without TAZ silencing were subjected to TAZ ChIP analysis using anti-TAZ or IgG control. The intronic region containing the TAZ/TEAD binding sequence, or a non-consensus sequence as control, was amplified by qPCR and normalized to the values obtained from input DNA (*p = 0.03; mean ± SEM; n=3).

(C) A dual-luciferase reporter assay in control and TAZ-silenced AML12 cells was conducted using the Ihh intron 1 region in (B), or a mutated version, together with 500 bp of flanking upstream and downstream sequence. (**p < 0.01, ***p < 0.0001; mean ± SEM; n=6).

(D) Immunoblot of Ihh in normal humam livers or those with steatosis or NASH.

(E) Immunoblot of Ihh in the livers of mice fed chow or FPC diet for 16 weeks.

(F) Relative expression of Ihh, Gli2, Gli3, and Opn mRNAs in the livers of mice fed chow or FPC diet for 16 weeks (*p < 0.04, **p < 0.0001; mean ± SEM; n=6).

(G) Relative expression of Ihh, Gli2, Gli3, and Opn mRNAs in the livers of mice fed the FPC diet for 16 weeks with or without TAZ silencing (*p < 0.05, **p < 0.002; mean ± SEM; n=10).

(H) Immunoblot of Ihh in the livers of mice fed the FPC diet for 16 weeks with or without TAZ silencing.

(I) OPN immunohistochemistry and quantification in the livers of mice fed the FPC diet for 16 weeks with or without TAZ silencing (*p < 0.0001; mean ± SEM; n=10); Bar, 200 μm.