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. 2017 Nov 9;7:15118. doi: 10.1038/s41598-017-15087-0

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The effect of PKA on the human long-NT and short-NT isoforms of Ca_v1.2. Schematics of human long-NT (a) and short-NT isoforms (e) indicating transmembrane repeats (RI-IV, grey bars) distinct exons 1a and 1b, and the amino (N) and carboxyl (C) terminals. (b,f) Immunoblots of the Ca_v1.2 channel protein indicated at ~240 kDa band probed with Anti-CaVpanα1 antibody. Anti-phosphoSer/Thr PKA substrate specific antibody (Anti-pS/pT) demonstrates phosphorylation after in vitro PKA phosphorylation (P). Subsequent dephosphorylation by PP2A (DP) removed the signal (C: control). Semi-quantitative western blot analysis demonstrated below (Rel. OD: relative optical density). Full-length blots are presented in Supplementary Figure 2. Representative single channel currents of the long-NT (c) and short-NT isoform (g) recorded at +150 mV in the absence (Con) and presence of PKA in the same patch (d,h) The current-voltage (I–V) relationship (right) and the mean ± SEM channel open probability (P_o) for currents recorded in the absence and presence of PKA (left). n: number of single channel patches, vertical scale bar indicates 2 pA, horizontal bar indicates 100 ms. *p < 0.05 PKA vs Con.