Figure 7

ROS production is increased in zebrafish embryos injected with NOX4 mRNA variants compared to NOX4^wt.

(A) Endogenous Nox4 was reduced by co-injecting embryos with a translation-blocking morpholino (nox4 atg MO), resulting in a 50% reduction of Nox4 compared to mock-injected controls. Gapdh was used as loading control. N = 3. (B) ROS production was assessed by imaging zebrafish embryos (red box) and quantifying the fluorescence intensity in the area marked by the blue box. (C) Quantification of fluorescence intensity shows a significant increase in the embryos co-injected with NOX4 variants mRNA and nox4 atg MO compared to NOX4^wt and nox4 atg MO The number of zebrafish embryos imaged in each experimental group is listed in Appendix Table S7. In total, 9–16 zebrafish embryos were used for each repeat. N = 3. (D) Representative images of injected embryos exposed to DCFH-DA (scale bars = 100 μm). Data information: DCFH-DA 2′,7′-dichlorofluorescein diacetate. N =  biological replicates. Values are mean ± SEM. For graph (A), the difference between two groups was determined by the Student’s t test. For graph (C), ordinary two-way ANOVA with Tukey’s multiple comparisons test was used to determine significant differences. Source data are available online for this figure.