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Fig. 2
VE-cadherin-TS is functional and under acto-myosin controlled tension. a Schematic representation of genetic cross to test functionality of VE-cadherin-TS. b Wild-type (top) and ve-cadherin mutant (bottom) phenotype at 2?dpf (arrowhead indicates cardiac oedema in mutant). Scale bar?=?1?mm. c Phenotypic scoring all embryos (n?=?430) collected from cross in a, showing 23% phenotypic mutants (n?=?49/217) in TS-negative population (n?=?217) and 0% (n?=?0/213) in TS-positive siblings (n?=?213). d Phenotypic scoring of n?=?286 embryos from a cross between a Tg(ve-cad:ve-cadTS)+/?, cdh5 ubs8?/? mutant and a non-transgenic cdh5 ubs8+/? animal. In the TS negative population 52% (n?=?76/146) displayed the mutant phenotype whilst there were no phenotypic mutants in the TS positive clutch (n?=?141). e Schematic representation of Y-27632 (ROCK inhibitor) acto-myosin inhibition. f Heatmap image of ratio-metric FRET values in junctions of 1%DMSO and Y-27632 (45??M/1%DMSO) treated embryos. Colors range from blue (?=?low FRET index/high tension) to red (?=?high FRET index/low tension). Scale bar?=?5??m. g Quantification of ratio-metric FRET values in junctions of 1 % DMSO (n?=?4) versus Y-27632 (45??M/1%DMSO) (n?=?5) treated embryos. Each data point represents a junctional region of interest (ROI), n?=?33 junctions selected from DMSO control embryos and n?=?40 selected in Y-27632 treated embryos. Treatment for 5?h prior to imaging. Error bars represent mean?±?s.d.; ****p?<?0.0001 from unpaired two-sided Mann?Whitney test. h Heatmap image of Teal lifetime values in junctions of 1%DMSO (top) and Y-27632 (bottom, 45??M/1%DMSO)- treated embryos. Colors range from blue (?=?low lifetime/low tension) to red (?=?high lifetime/high tension). Scale bar?=?5?m. i Quantification of Teal lifetime values (nano-sec) in junctions of 1 % DMSO (n?=?4) versus Y-27632 (45??M/1%DMSO) (n?=?4) treated embryos. Each data point represents a single junctional ROI, n?=?25 junctions segmented from DMSO control embryos and n?=?33 from Y-27632 treated embryos. Treatment for 5?h prior to imaging. Error bars represent mean?±?s.d.; **p?=?0.0060 from unpaired two-sided t test