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. 2008 Feb 7;36(5):1690–1702. doi: 10.1093/nar/gkn009

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© 2008 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 7. — All five zinc finger motifs are required for optimal binding of Glis3 to the consensus Glis-BS. (A) A schematic view of His₆-Glis3(ZFD)1–5m mutants in which the first Cys in each zinc finger motif is mutated to Ala (left panel). PAGE analysis of His₆-Glis3(ZFD) proteins used in EMSA (upper panel). (B) EMSA using His₆-Glis3(ZFD) and His₆-Glis3(ZFD)1–5m mutant proteins and ³²P-labeled consensus Glis-BS. The position of the Glis3/Glis-BS complex is indicated by the arrow. (C) The ability of VP16-Glis3 and VP16-Glis3(ZFD)1–5m mutants to induce (Glis-BS)₆-dependent transactivation of the LUC reporter gene was examined in HEK293T cells. (D) The ability of VP16-Glis3 and mutants VP16-Glis3(ΔC625) and VP16-Glis3(ΔN532), encoding a C- and N-terminal deletion mutant of Glis3 as indicated in the lower panel, to induce (Glis-BS)₆-dependent transactivation of the LUC reporter gene was examined. The relative luciferase activity was calculated and plotted. Numbers indicate amount of plasmid (μg) transfected.