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, cygb2^801a embryos, wt treated with 500 μM cPTIO and DMSO as control and cygb2^801a 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 cygb^801a 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 cygb2^pt801a 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 cygb2^pt801a 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.