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. 2021 Jan 11;11:591773. doi: 10.3389/fphar.2020.591773

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Copyright © 2021 Bollmann, Werner, Jaron, Bruns, Wache, Runte, Boknik, Kirchhefer, Müller, Buchwalow, Rothemund, Neumann and Gergs.

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) and the copyright owner(s) 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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Generation of transgenic mice. (A) Transgenic construct: the cDNA of bovine PP2Cβ was overexpressed under the control of the α-myosin heavy chain (α-MHC) promoter, which conveys cardiac myocyte selectivity of gene expression. (B) Western blot experiments confirmed a successful overexpression of PP2Cβ protein in PP2C-TG hearts (TG) compared to WT. Molecular weight standards are indicted in kilo Daltons (kDa) on the left hand side of the gel. Cardiac calsequestrin (CSQ) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) detection on the blotting membrane indicates equal protein loading. (C) The putative activity of PP2C in cardiac homogenates was determined using the artificial substrate p-nitrophenyl phosphate (p-NPP) in presence of Mn²⁺. (D) Quantitative analysis of Western blot experiments revealed unchanged cardiac protein expression of CSQ, sarco/endoplasmic reticulum Ca²⁺ ATPase (SERCA), phospholamban (PLB), and troponin inhibitor (TnI) in PP2C-TG hearts compared to WT. Moreover, PP2C was nearly tenfold overexpressed in PP2C-TG hearts. ★p < 0.05 vs. WT; ⁺ p < 0.05 vs. basal.