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. 2017 May 10;9:8. doi: 10.3389/fnsyn.2017.00008

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Copyright © 2017 Nair, Patil, Tiron, Kanhema, Panja, Schiro, Parobczak, Wilczynski and Bramham.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Enhanced Arc SUMOylation in the cytoskeletal fraction during LTP in vivo. (A) Immunoblot characterization of dentate gyrus subcellular fractions. Ipsilateral HFS-treated (Ipsi), contralateral control (Contra), and Naïve. Vimentin and histone 1 were used as markers of the cytoskeletal and nuclear fractions, respectively. Tissue was collected 1 h post-HFS and fractions collected from four dentate gyri were pooled. A 65 kDa Arc immunoreactive band (indicated by an arrow) was detected in the cytoskeletal fraction. (B) Bidirectional co-immunoprecipitation using anti-SUMO1 and anti-Arc antibodies was performed in dentate gyrus cytoskeletal fractions. SUMO1ylated Arc was detected at 65 kDa. Representative blots based on three independent biological replicates. Sameples from two dentate gyrus were pooled. Note the absence of unmodified Arc in the SUMO1 pellet. (C) Arc immunoprecipitation was performed in the cytoskeletal fraction from HFS-treated and control dentate gyrus. (D) Arc immunoblot analysis of cytoskeletal fraction. Bar graphs shows increase in Arc expression and enhanced Arc SUMOylation based on the ratio of upper/lower Arc bands in the dentate gyrus cytoskeletal fraction at 1 h post-HFS relative to contralateral control. n = 5; Student’s t-test, *P < 0.05. Representative Arc immunoblots on right. Arrow indicates 65 kDa SUMOylated Arc.