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. 2009 Feb 23;184(4):491–500. doi: 10.1083/jcb.200807175

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© 2009 Conforti et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 1. — Overexpression of ΔN16Wld^S and ATX3Wld^S in Tg mice. (A and B) Western blots of ΔN16Wld^S and ATX3Wld^S brains probed with Wld18. ΔN16Wld^S lines 1 and 2 alone express a protein slightly smaller than Wld^S. The 43-kD ATX3Wld^S band matches that in Wld^S. (C) Brain Western blots of total homogenate and nuclear and cytoplasmic fractions of Wld^S, ΔN16Wld^S (line 1), and ATX3Wld^S (line 6). The graph shows integrated band intensities of nuclear and cytoplasmic fractions normalized to H1 and β-actin, respectively. These normalized figures were then expressed as a percentage of the total homogenate signal and normalized to the same respective markers (mean ± SD; n = 3). Statistical analysis was performed on the nuclear versus supernatant ratio using a Mann-Whitney test followed by a Bonferroni post-hoc test. (D) Immunofluorescence of lumbar spinal cord sections with Wld18 (red) and DAPI. Motor and interneuron nuclear signals in Tg ΔN16Wld^S (i–iv) and ATX3Wld^S (v–viii) show similar strength and distribution as Wld^S heterozygotes. Identical laser intensities and camera settings were used for each image. (E and F) Transgene products are enzymatically active. Nmnat activity is very significantly increased compared with wild-type (WT) brains in hemizygotes and homozygotes of all expressing Tg lines as well as Wld^S heterozygotes. Mean ± SD; *, P < 0.05; **, P < 0.01; ***, P < 0.001. Bars, 10 µm.