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. 2015 Mar 1;29(5):501–512. doi: 10.1101/gad.258483.115

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© 2015 Murn et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 1. — Evolutionary conservation and expression of Unkempt. (A) Sketch of the human Unkempt protein showing the relative positions of the six tandem CCCH zinc finger motifs and a RING finger domain. Bar, 100 amino acids. (B) Unkempt protein is conserved across metazoans. The cladogram shows the percentage of amino acid identities of Unkempt orthologs found in each of the indicated species compared with the full-length human ortholog. Note that no orthologs of Unkempt are found in nonmetazoan species. Evolutionary relationships of animals shown are based on Srivastava et al. (2010). (Adapted by permission from Macmillan Publishers Ltd., © 2010.) RefSeq protein accession numbers of all of the indicated Unkempt orthologs are listed in Supplemental Figure S1A. (C) Detection of Unkempt (UNK) in continuous cell lines by immunoblotting. (D,H) Depletion of endogenous Unkempt in SH-SY5Y cells detected by immunoblotting (D) and immunofluorescence (H) using Unkempt-specific antibodies. Bar, 10 μm. (E) Immunohistochemistry of mouse E15 whole embryo sagittal sections revealing highly enriched expression of Unkempt in the CNS. The location of the CNS is highlighted by the expression of the neuronal marker NeuN. See also Supplemental Figure S1D. (F) Expression of Unkempt in the cortex of E15 mouse embryos. The cortical wall was probed with antibodies against Unkempt (green) and neuronal marker Tuj-1 (red) and counterstained with DAPI to label the nuclei (blue). The highlighted region (orange rectangle) is shown magnified (middle), along with a close-up of the ventricular zone (right), together indicating a pervasive expression of Unkempt throughout the cortical wall. Bars, 10 μm. (G) Immunocytochemistry of dissociated E15 neurons in vitro showing the expression of Unkempt and the neuronal marker Tuj-1. Bar, 10 μm.