FIGURE SUMMARY
Title

ZNF408 is mutated in familial exudative vitreoretinopathy and is crucial for the development of zebrafish retinal vasculature

Authors
Collin, R.W., Nikopoulos, K., Dona, M., Gilissen, C., Hoischen, A., Boonstra, F.N., Poulter, J.A., Kondo, H., Berger, W., Toomes, C., Tahira, T., Mohn, L.R., Blokland, E.A., Hetterschijt, L., Ali, M., Groothuismink, J.M., Duijkers, L., Inglehearn, C.F., Sollfrank, L., Strom, T.M., Uchio, E., van Nouhuys, C.E., Kremer, H., Veltman, J.A., van Wijk, E., and Cremers, F.P.
Source
Full text @ Proc. Natl. Acad. Sci. USA

Immunocytochemical analysis of COS-1 cells transiently transfected with constructs encoding HA-tagged WT and mutant (p.Ser126Asn, p.His455Tyr) ZNF408 proteins. DAPI stains cell nuclei, whereas the anti-HA antibody stains the ZNF408 fusion proteins (indicated by arrows). (Right) Merged pictures (DAPI in blue, and HA-tagged ZNF408 proteins in red). Representative examples are shown for each transfection, indicating full nuclear localization for the WT and p.Ser126Asn ZNF408 proteins, but with the p.His455Tyr mutant ZNF408 confined mainly to the cytoplasm (each indicated by arrows).

Cotransfection analysis of WT and p.His455Tyr mutant ZNF408. (A–C) Immunocytochemical analysis of transiently transfected HA-tagged WT ZNF408 proteins. Representative examples show WT ZNF408 expression inside the cell nuclei (arrows). Images are presented as DAPI (staining cell nuclei) (A), anti-HA (detecting HA-tagged ZNF408 proteins) (B), and merged pictures (DAPI in blue, HA-tagged ZNF408 in red) (C). (D–F) Fluorescence analysis of transiently transfected eCFP-p.His455Tyr mutant ZNF408 proteins. Mutant ZNF408 is located mainly outside the cell nuclei (indicated by arrows). Images are presented as DAPI (staining cell nuclei) (D), eCFP (detecting eCFP-ZNF408 mutant proteins) (E), and merged pictures (DAPI in blue, eCFP-tagged p.His455Tyr ZNF408 in green) (F). (G–J) Combined analysis of COS-1 cells cotransfected with HA-tagged WT ZNF408 as well as eCFP-p.His455Tyr mutant ZNF408 proteins. In cells that express both the mutant and the WT ZNF408, the WT ZNF408 is also retained in the cytoplasm (I and J). Images are presented as DAPI (staining cell nuclei) (G), eCFP (detecting eCFP-p.His455Tyr ZNF408 mutant proteins (H), anti-HA (detecting HA-tagged WT ZNF408 proteins) (I), and merged pictures (DAPI in blue, HA-tagged WT ZNF408 in red, eCFP-p.His455Tyr ZNF408 in green) (J).

Cotransfection analysis of WT and p.His455Tyr mutant ZNF408. (A–C) Immunocytochemical analysis of transiently transfected HA-tagged WT ZNF408 proteins. Representative examples show WT ZNF408 expression inside the cell nuclei (arrows). Images are presented as DAPI (staining cell nuclei) (A), anti-HA (detecting HA-tagged ZNF408 proteins) (B), and merged pictures (DAPI in blue, HA-tagged ZNF408 in red) (C). (D–F) Fluorescence analysis of transiently transfected eCFP-p.His455Tyr mutant ZNF408 proteins. Mutant ZNF408 is located mainly outside the cell nuclei (indicated by arrows). Images are presented as DAPI (staining cell nuclei) (D), eCFP (detecting eCFP-ZNF408 mutant proteins) (E), and merged pictures (DAPI in blue, eCFP-tagged p.His455Tyr ZNF408 in green) (F). (G–J) Combined analysis of COS-1 cells cotransfected with HA-tagged WT ZNF408 as well as eCFP-p.His455Tyr mutant ZNF408 proteins. In cells that express both the mutant and the WT ZNF408, the WT ZNF408 is also retained in the cytoplasm (I and J). Images are presented as DAPI (staining cell nuclei) (G), eCFP (detecting eCFP-p.His455Tyr ZNF408 mutant proteins (H), anti-HA (detecting HA-tagged WT ZNF408 proteins) (I), and merged pictures (DAPI in blue, HA-tagged WT ZNF408 in red, eCFP-p.His455Tyr ZNF408 in green) (J).

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Proc. Natl. Acad. Sci. USA