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. 2016 Jul 27;64(11):1938–1961. doi: 10.1002/glia.23034

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© 2016 The Authors. Glia Published by Wiley Periodicals, Inc.

This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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Immunoreactivity of 9F5 was not affected by deglycosylation of rat GPNMB protein. A: Extracts of rat type 1 MG were treated with or without N‐glycosidase F (PNGase F) overnight at 37°C, and they were then subjected to immunoblotting with 9F5 (a), anti‐GPNMB (b), or ED1 (c) antibodies. As reported previously for murine dendritic cells (Shikano et al., 2001), full‐length GPNMB (119.6 and 94.9 kDa) in type 1 MG shifted to 68.3 and 79.8 kDa after PNGase F treatment (Ab). B: Extracts of type 1 MG were treated with or without sialidase and O‐glycosidase overnight at 37°C, and they were then subjected to immunoblotting with 9F5 (a), anti‐GPNMB (b), or ED1 (c) antibodies. Immunoreactivity of ED1, an antibody that reacts with sialic acid on rat CD68, disappeared after sialidase treatment (Bc).