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. 2008 Aug 11;182(3):531–542. doi: 10.1083/jcb.200711151

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© 2008 Ju 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. — Localization of human Cosmc and T-synthase. (A–F) Immunofluorescent staining. CHO K1 cells cultured on chambered slides were transiently transfected with Cosmc-HA or with T-synthase–HPC4 and stained with rat anti-HA IgG1 (green) and rabbit anti-calnexin IgG (red; A–C; bars, 4 μm) or with mouse anti-HPC4 (green) and rabbit anti–α-ManII (red; D–F; bars, 8 μm). (G and H) Sucrose gradient subcellular fractionation. 293T cells transiently transfected with HPC4-Cosmc were harvested and homogenized. The postnuclear supernatant (PNS) was loaded onto a sucrose gradient. After ultracentrifugation, 16 fractions (∼0.6 ml/fraction) were collected and measured for both T-synthase and β4–Gal-T activity (G) and analyzed on Western Blot with anti-HPC4 and anti-KDEL (H).