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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 9. — Glis3(NDH1) localized to the nucleus, but lost its transactivation activity. (A) Schematic view of Glis3(Δ155) and Glis3(NDH1). ZDF and AD are indicated. An insertion of C (arrow head) in the Glis3 gene (NDH1) is linked to the most severe type of NDH. This insertion results in a frame shift at 779 and a stop codon at 862 (779FS862STOP) as indicated (shaded box). (B) Glis3(NDH1) localizes to the nucleus. HEK293T cells were transfected with p3×FLAG-CMV-Glis3(NDH1) and subcellular localization was examined by confocal microscopy with anti-FLAG M2 and Alexa Fluor 594 antibodies; nuclei were identified by DAPI staining. (C) Glis3(NDH1) lacks transcriptional activity. HEK293T cells were co-transfected with p3×FLAG-CMV-Glis3(ΔN155), p3×FLAG-CMV-Glis3(NDH1), pCMVβ and (Glis-BS)₆-LUC as indicated. After 30 h, cells were assayed for luciferase (LUC) and β-galactosidase activity. The relative LUC reporter activity was calculated and plotted. The expression of Glis3(ΔN155) and Glis3(NDH1) proteins was examined by western blot analysis using anti-FLAG antibody (lower panel).