Figure 7

Zebrafish were treated with 40 µM SFN or 1 µM tBHQ during the hatching and protruding-mouth stages for 6 h and incubated with BioGee for 2 h before fixation 24 h after treatment. BioGee protein conjugates were labeled in situ using IHC. Confocal microscopy was used to acquire images of the liver using a 40× objective. (A) Liver mean fluorescence intensity (FI) of BioGee protein conjugates was determined via image analysis. Representative images of the zebrafish liver treated during the (B) hatching/protruding-mouth and (C) protruding-mouth/larval stage are shown where the liver is outlined in yellow, TRITC (purple) represents BioGee, and DAPI (blue) represents nuclei. The liver is not present (n.p.) at the stage visualized. Statistics 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) among groups. Scale bar = 50 µm.