Figure 6

Figure 6

Zebrafish were treated with 40 µM SFN or 1 µM tBHQ for 6 h during the pharyngula, hatching, and protruding-mouth stages and incubated with BioGee for 2 h before fixation 24 h after the start of the exposure. BioGee protein conjugates were labeled in situ using IHC. Confocal microscopy was used to acquire Z-stacks of the islet. (A) Islet volume and (B) islet mean fluorescence intensity (FI) of BioGee. Representative images at the (C) pharyngula (start of exposure)/hatching (BioGee labeling and fixation), (D) hatching/protruding-mouth, and (E) protruding-mouth/larval stages are shown. Images are max intensity projections of the Z-stack the islet (outlined in yellow), where FITC (green) represents β-cells, TRITC (purple) represents BioGee protein conjugates, and DAPI (blue) represents nuclei. Statistical significance was assessed via two-way ANOVA followed by Fisher’s LSD post hoc test. n = 7–9 fish. Letters indicate significant differences (p ≤ 0.05) among groups. Scale bar = 25 µm.