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Figure 4
Zebrafish were treated with 40 µM SFN or 1 µM tBHQ during the pharyngula, hatching, and protruding-mouth stages for 6 h and then fixed, and Nrf2a protein was labeled using immunohistochemistry (IHC). Colocalization analysis was performed on 40× confocal images of the liver, and a representative image of the pancreatic islet was taken from the Z-stack. Pearson’s R coefficients were converted to normally distributed Z-scores, shown as means ± SEMs, where higher Z-scores indicate greater colocalization between Nrf2a protein and nuclear DAPI staining within the region of interest. Significance was assessed with a two-way or three-way ANOVA followed by Fisher’s LSD post hoc test.