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. 2020 Apr 22;295(22):7789–7798. doi: 10.1074/jbc.RA120.013444

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© 2020 Kobayashi et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 1. — Isolation of human PrP^C from KI mouse brains. A, schematic of GPI-AP composition. Core and side-chain structures of mammalian GPI-APs are shown. The cleavage site of PI-PLC is indicated by an arrow. Glycan symbols follow the symbol nomenclature for glycans (44). B, PI-PLC treatment of the mouse medulla oblongata. Medulla oblongata homogenates obtained from mice expressing human PrP^C were treated with PI-PLC, and then free GPI-GalNAc was detected by Western blotting. GAPDH was used as a loading control. C, isolation of human PrP^C from KI mouse brains. The lipid portion of the GPI anchor of human PrP^C was removed by PI-PLC, followed by immunoprecipitation with an anti-prion antibody. N-glycans on isolated PrP^C were removed by treatment with PNG-F. Bands corresponding to PNG-F and human PrP^C are indicated by arrows.