Hearts were isolated from male C57BL/6J male mice (~20–25 g) (Charles River JAXTM stock number 000664) or from WT and PLM3SA knock-in mice18 (link). This investigation complied with all the relevant ethical regulations for animal testing and research: UK Home Office Guidance on the Operation of the Animals (Scientific Procedures) Act, 1986.
PLM3SA knock-in mice were backcrossed with C57BL/6J mice (Charles River, UK) for >5 generations and were generated by heterozygous pair mating. The PLM3SA mouse expresses a non-phosphorylatable form of PLM in which Ser 63, 68, and 69 have been mutated to alanine and exhibits a Na/K ATPase that is unresponsive to kinase regulation and hence shows chronically elevated Nai15 (link). Unless otherwise stated, littermates were used as the appropriate wild-type controls (PLMWT). Animals were kept under pathogen-free conditions, 12-h light–dark cycle, controlled humidity (~40%), temperature (20–22 °C), and fed chow and water ad libitum. All animals used in studies were male. For pharmacologically induced acute Nai elevation studies, 6-week-old C57BL/6J male mice (~25 g body weight) were purchased from Charles River (UK). Myocardial hypertrophy was induced in 6-week-old C57BL/6J mice (20–22 g) (Charles River, UK). For cellular compartmentation of the 23Na TQF NMR signal experiment, Male Wistar rats (250 g) were purchased from Charles River, UK.
PLM3SA knock-in mice were backcrossed with C57BL/6J mice (Charles River, UK) for >5 generations and were generated by heterozygous pair mating. The PLM3SA mouse expresses a non-phosphorylatable form of PLM in which Ser 63, 68, and 69 have been mutated to alanine and exhibits a Na/K ATPase that is unresponsive to kinase regulation and hence shows chronically elevated Nai15 (link). Unless otherwise stated, littermates were used as the appropriate wild-type controls (PLMWT). Animals were kept under pathogen-free conditions, 12-h light–dark cycle, controlled humidity (~40%), temperature (20–22 °C), and fed chow and water ad libitum. All animals used in studies were male. For pharmacologically induced acute Nai elevation studies, 6-week-old C57BL/6J male mice (~25 g body weight) were purchased from Charles River (UK). Myocardial hypertrophy was induced in 6-week-old C57BL/6J mice (20–22 g) (Charles River, UK). For cellular compartmentation of the 23Na TQF NMR signal experiment, Male Wistar rats (250 g) were purchased from Charles River, UK.
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