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Fig. 4

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ZDB-IMAGE-231218-5
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Figures for Rochon et al., 2023
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Fig. 4 Cygb2 regulates Nos2b-NO-sGC signaling in KV cilia function and cardiac laterality determination.

a Beads were injected into the KV between 8–10 somites and velocity was measured over time. Representative bead tracks of the KV comparing wt, cygb2801a embryos, wt treated with 500 μM cPTIO and DMSO as control and cygb2801a treated with 250 μM DETA/NO and NaOH/PBS as control (n = 4 embryos per treatment). The dot indicates the end of the bead track after ten seconds of tracking. Scale bar = 20 μm. Boundaries of the KV are demarcated by a white dashed line. b Quantification of bead velocities (n = 20–35 beads over 4 embryos). Means are ± SD. One-way Anova, ***P = 0.003; ****P < 0.0001. c Analysis of cardiac laterality defects represented as a percentage of embryos with the correct heart looping (on the left) analyzed at 2 dpf. Wt and cygb801a treated with 250 µM DETA/NO or 500 μM cPTIO at the end of the blastula period. Means are ± SD, (n = 4). Student’s t test, two-tailed, ns = not significant, *P < 0.05, **P < 0.01. d Experimental schematic of the developmental time course drug treatment with 250 µM DETA/NO. Panel generated with BioRender.com. e Percentage of embryos with right, midline or left heart orientation following treatment with DETA/NO beginning at different developmental stages. The total number of embryos analyzed is shown in red above the graph. The Chi-squared test was used to determine statistical significance. f Quantification of the percentage of embryos with left hearts comparing wt to cygb2pt801a with and without treatment of 80 µM cinaciguat. Means are ± SD, (n = 4–6). Student’s t test, two-tailed, ns = not significant, ** P < 0.01. g Quantification of the percentage of embryos with left heart comparing wt to cygb2pt801a injected with nos2b mRNA compared to uninjected controls. Means are ± SD, (n = 6–7). Student’s t test, two-tailed, *P < 0.05, ns = not significant. In (c, f, g): each sample represent an independent experiment consisting of 50 embryos. Source data are provided as source data file.

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