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. 2019 Dec 9;8(12):bio046474. doi: 10.1242/bio.046474

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© 2019. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 1. — Generation of tnnt2a mutant zebrafish and phenotype analyses. (A) Graphical representation of the target site of tnnt2a for CRISPR/Cas9 gRNA and altered amino acid sequence of Tnnt2a of mutant zebrafish. (B) Genotype sequencing results of tnnt2a^+/+ and tnnt2a^−/− zebrafish. (C) Representative images of atrial (A) and ventricular (V) enlargement, and pericardial effusion in 72 hpf tnnt2a^−/− zebrafish. Scale bars: 150 µm. (D–G) Statistics for the heart rate, atrial and ventricular diameter, and pericardium size in wild-type and tnnt2a^−/− zebrafish; ****P<0.0001; n=15 per group. Statistical differences were evaluated using two-tailed unpaired t-tests (D–F) or unpaired t-test with Welch's correction (G). (H,I) Fluorescent images and pseudo-colours of electrocardiac rhythm in Tg(bactin2:GCamp6s; tnnt2a^+/+) and Tg(bactin2:GCamp6s; tnnt2a^−/−) zebrafish; arrows represent increased Ca²⁺ flux in cardiomyocytes. The direction of Ca²⁺ flux was from venous to arterial end both in tnnt2a^+/+ and tnnt2a^−/−. Scale bars: 100 µm.