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Fig. 5
Calcium signaling maintains lower VE-cadherin tension during artery maturation. a Heatmap image showing average intensity projection of fli1a-driven GCaMP5G expression (indicating calcium) in the DA at 2?dpf (left) and 4?dpf (right). Colors range from black (?=?low calcium) to white (?=?high calcium). Scale bar?=?10??m. b Increase in fluorescence intensity index (?=?fli1a driven GCaMP5G/ fli1a-driven RFP) in the dorsal wall of the DA of n?=?9 embryos imaged at 2 and 4?dpf. **p?=?0.0012, from paired t test. c Junctional morphology of ECs in the DA (Venus, grey) of a 0.5% DMSO, a 15??M/0.5% DMSO Nifedipine and a 25??M/0.5% DMSO Nifedipine treated embryo at 4?dpf. Treatment from 2 to 4?dpf. Scale bar?=?10??m. d Ratio-metric FRET values in junctions of 0.5% DMSO (n?=?42 junctional ROIs from n?=?5 embryos), 15??M Nifedipine (n?=?54 junctional ROIs from n?=?6 embryos) and 25??M Nifedipine (n?=?59 junctional ROIs from n?=?6 embryos) treated embryos. Treatment was from 2 to 4?dpf. Error bars represent mean?±?s.d.; DMSO ? 15??M Nifedipine **p?=?0.0068, DMSO ? 25??M Nifedipine ****p?<?0.0001, from unpaired two-sided Mann?Whitney test.e Junctional linearity index in 0.5% DMSO treated controls (n?=?5) versus 25??M Nifedipine-treated embryos (n?=?6). Each data point represents a junction, n?=?18 control junctions measured and n?=?18 junctions from Nifedipine-treated embryos. Error bars represent mean?±?s.d.; not significant (ns) p?=?0.8147, from unpaired two-sided Mann?Whitney test. f Ratio-metric FRET values quantified from 1% DMSO-treated controls (n?=?39 junctional ROIs from n?=?6 embryos), BAPTA-AM-treated (100??M for 1?h, n?=?58 junctional ROIs from n?=?8 embryos), Nifedipine-treated (50??M for 30?min, n?=?27 junctional ROIs from n?=?5 embryos) and Nifedipine?+?Y-27632-treated embryos (n?=?47 junctional ROIs from n?=?8 embryos). Error bars represent mean?±?s.d.; 1% DMSO ? 100??M BAPTA-AM *p?=?0.0498; 1% DMSO ? 50??M Nifedipine ****p?<?0.0001; 50??M Nifedipine - 50??M Nifedipine?+?50??M Y-27632 ***p?=?0.0008, from unpaired two-sided t test