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. 2018 Sep 3;8:13156. doi: 10.1038/s41598-018-31584-2

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© The Author(s) 2018

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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Effects of pre-incubation with the Ca²⁺-calmodulin‐dependent kinase type II (CaMKII) inhibitor KN-93 (1 μM) on the TSA- (1 μM) and MC1568- (1 μM) induced reduction of I_Ca. (A) Standard current-voltage (I-V) relationship of I_Ca after 30 min incubation with KN-93 (n = 5 PPN neurons). (B) Representative I_Ca recorded using 100 msec long depolarizing square steps from a holding potential of −50 mV to 0 mV after 30 min incubation with KN-93 (1 μM; black record) and after acute bath application TSA (20 min, 1 μM; green record). (B) Percent reduction in I_Ca amplitude in a group of PPN cells (n = 8) recorded after acute bath application TSA (20 min, 1 μM; black bar), and following pre-incubation with KN-93 (1 μM; n = 7; green bar). **p < 0.01 significant reduction of I_Ca by acute bath application of TSA (Paired Student’s t-test; t = −3.77; df = 7; p = 0.007), or KN93 + TSA (paired Student’s t-test; t = −3.96; df = 6; p = 0.007). Note the statistically significant KN-93-dependent blunting effect of the TSA-induced reduction of I_Ca (One-way ANOVA; F_(1,13) = 6.3; p < 0.05; post-hoc Bonferroni’s t-test, t = 2.51, p = 0.03). (C) Percent reduction in I_Ca amplitude in a group of PPN cells (n = 6) recorded after acute bath application MC1568 (20 min, 1 μM; solid grey bar), and following pre-incubation with KN-93 (1 μM; n = 6; dashed grey bar). *p < 0.05 significant reduction of I_Ca by acute bath application of MC1568 (n = 6 PPN cells; paired Student’s t-test, t = 3.46; df = 5, p = 0.018), or KN93 + MC1568 (n = 6 PPN cells; paired Student’s t-test, t = −3.75; df = 5, p = 0.013). Note the statistically significant blunting effect of KN-93 on the mean I_Ca reduction by MC1568 (One-way ANOVA; F_{(1, 10)} = 9.24; p = 0.013; post-hoc Bonferroni t-test; t = 3.04; p < 0.05).