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Fig. 5

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ZDB-IMAGE-211230-47
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Figures for Kamel et al., 2021
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Figure Caption

Fig. 5 A Schematic representation of Ca2+ transient parameters showing Ca2+ transient upstroke time, Ca2+ transient recovery time, Ca2+ transient amplitude, and diastolic Ca2+ levels. B Representative image of an isolated ventricular cardiomyocyte and a Ca2+ trace from adult ventricular cardiomyocyte of wild type and plna R14del mutants. Intracellular Ca2+ parameters were measured in adult ventricular cardiomyocytes of wild type (WT) and plna R14del, including C diastolic Ca2+ levels (mean ± SEM, *p = 0.05, WT n = 4, plna R14del n = 5), D systolic Ca2+ levels, E Calcium transient duration at 20%, 50, and 80% (mean ± SEM, CaD20: p = 0.026 and CaD50: p = 0.005, WT n = 4, plna R14del n = 5), and F Tau of Ca2+ decay (mean ± SEM, *p = 0.041, WT n = 4, plna R14del n = 5). All measurements were performed in three experimental replicates. G DNA construct and sensor dynamics of GCaMP6f (left panel). GCaMP6f was placed under the control of the myl7 promoter to restrict its expression to the heart. The Gal4FF-UAS system amplifies its expression. GCaMP6f consists of a circularly permutated enhanced green fluorescence protein (cpEGFP) fused to calmodulin (CaM) and the M13 peptide. When intracellular calcium (Ca2+) rises, CaM binds to M13, causing increased brightness of cpEGFP. Using a high-speed epifluorescence microscope, movies of 3 dpf non-contracting GCaMP6f embryonic hearts were recorded. Several intracellular Ca2+ parameters were measured, including H Frequency of Ca2+ transients, I Ca2+ transient recovery time, J normalized Ca2+ transient amplitude (mean ± SEM, *p = 0.0129, WT n = 10, plna R14del n = 12), K normalized diastolic Ca2+ levels, and L Tau of Ca2+ decay in wild-type (WT) and plna R14del embryonic zebrafish (mean ± SEM, *p = 0.0498, WT n = 10, plna R14del n = 12). All measurements were performed in three experimental replicates. Wild type is highlighted in black and plna R14del in red. Statistical test: unpaired Student’s t-test. nM nanomolar, ms milliseconds, CaM calmodulin, UAS upstream activation sequence, Ca2+ calcium. Source data are provided as a Source Data file.

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