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. 2017 Oct 4;9:13. doi: 10.3389/fnsyn.2017.00013

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Copyright © 2017 Meese, Cepeda, Gahlen, Adams, Ficner, Ricci, Heller, Reisinger and Herget.

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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Immunoprecipitation and western blot show interaction of otoferlin with CaMKIIδ. (A–C) Two HA-tagged mouse otoferlin fragments, C₂ABC (aa 1–632 in NP_001093865; 70 kDa) and C₂DEF (aa 933–1920; 114 kDa) were co-transfected with mcherry-tagged mouse CaMKIIδ into HEK293 cells. Transfections were performed either with otoferlin C₂ABC and CaMKIIδ (A, Input Lane 1 and 2), otoferlin C₂DEF and CaMKIIδ (B, Input Lane 1 and 2) or in the presence of both C₂ABC and C₂DEF fragments and CaMKIIδ (C, Input Lane 1 and 2). Co-immunoprecipitations of C₂ABC-HA and C₂DEF-HA were conducted from HEK293 cell lysates using anti-HA antibodies. CaMKIIδ-mcherry was detected in the eluate using an anti-RFP (red fluorescent protein) antibody (A–C, Lane 3), indicating that CaMKIIδ co-precipitated with recombinant otoferlin fragments.