Fig. 4.

Zebrafish were treated with 7 μM DMF during the pharyngula, hatching, and protruding-mouth stage for 6 hours and then immediately fixed and Nrf2a protein was labeled via Immunohistochemistry (IHC). Z-stacks were taken of the entire endocrine pancreas using a confocal microscope under a 40x objective. A) Islet volume and B) islet mean fluorescence intensity (FI) of Nrf2a protein was determined via a batch analysis workflow was using Nikon NIS elements software. Representative images of the wild type (WT) zebrafish at the C) pharyngula and D) protruding-mouth stage are shown to demonstrate differences with DMF treatment. Images are max intensity projection of the z stack the pancreatic islet (circle in yellow) where FITC (green) represents the beta cells, TRITC (red) represents Nrf2a protein, and DAPI (blue) represents nuclei. Calculations were performed using a two-way ANOVA followed by Fisher’s LSD post-hoc test. N = 6–12 fish. Different letters indicate significant differences (p ≤ 0.05) between treatment, genotype, and time point.