Fig. 2.

Exposure paradigm and data analysis pipeline for the larval chemomotor response assay. (A) Naïve larval zebrafish were cultured from 6–120 hpf, then exposed to test chemicals at 120 hpf to evaluate acute locomotor behavioral effects of various flavonoids and neuropharmacologicals across a concentration range. (B) Example of a 5-hour time-course of locomotor activity following exposure to (S)-equol across a broad concentration range (1–50 µM) in 0.1% DMSO. (C) Locomotor activity was binned into 30 min time-bins across the 5-hour exposure. (D) Within each 30 min time-bin, locomotor activity was normalized to the relative control (percent activity). Heat maps of locomotor activity were generated using binned and normalized data to comparatively evaluate locomotor responses of chemicals sets. *Significance determined using two-way repeated measures ANOVA with Tukey’s post-hoc, p ≤ 0.05, N = 16 animals per concentration. Data transformed using Box-Cox power transformation for normality prior to analysis. Data are reported as mean ± SD for normalized activity relative to the control within each time group. Summary data and statistics (time-course, binned, and normalized plots, 95% CI, N-values and p-values) for all tested flavonoids and neuropharmacologicals are provided in Supplementary Materials.