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. 2019 May 7;6(2):ENEURO.0434-18.2019. doi: 10.1523/ENEURO.0434-18.2019

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Copyright © 2019 Gutierrez et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 7. — Csmd2 is found in synaptosomal and postsynaptic fractions. A, Membrane fractionation of P30 mouse forebrain lysate using a Percoll gradient. Representative membrane fractions are shown at top. Equal amounts of protein from each fraction were analyzed by Western blot analysis using three different α-Csmd2 antibodies and an α-PSD-95 antibody. All three Csmd2 antibodies detected Csmd2 enriched in the synaptosome-containing fractions, along with PSD-95. B, Preparation of crude synaptosomes showed a similar enrichment of Csmd2 and PSD-95 in the syaptosomal fraction (P2 pellet) compared to the soluble fraction (S2). Further extraction of P2 with Triton X-100 showed Csmd2 enriched in the PSD fraction (TxP pellet) with PSD-95, compared to the Triton X-100-soluble fraction (TxS). The Novus antibody detected full-length Csmd2 and a smaller band of unknown identity at ∼150 kDa.