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. 2015 Jul 2;14(10):2550–2563. doi: 10.1074/mcp.M114.041533

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© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

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Fig. 8. — BACE1 increases shedding of PLXDC2, PTPRN2 and ENPP5. Shown are immunoblots of the indicated proteins. Human embryonic kidney HEK293T cells were stably transfected with HA and FLAG epitope-tagged proteins (HA-PLXDC2-FLAG (A), HA-PTPRN2-FLAG (B), HA-ENPP5-FLAG (C) or the empty pcDNA3.1 vector as control (CON). Additionally, the stable cells were transiently transfected with BACE1 or empty peak12 vector as indicated. Additionally the cells were treated with either the BACE1 inhibitor C3 or DMSO. The recombinant full-length (fl) proteins PLXDC2, PTPRN2 and ENPP5 were detected in the cell lysate (lys) using the N-terminally binding HA antibody and the C-terminally binding FLAG antibody (indicated by arrowheads). The soluble (s) ectodomains sPLXDC2, sPTPRN2, sENPP5 were immunoprecipitated from the conditioned medium with the HA.7 monoclonal antibody and detected using the HA monoclonal antibody 3F10 (indicated by arrowheads). Calnexin served as a loading control. BACE1 was detected by 3D5 antibody. Shown are representative immunoblots from three independent experiments.