Figure 6.

Developmental and behavioral assays and media and tissue concentrations in zebrafish exposed to GenX Free Acid diluted in DI water. (A) Developmental toxicity scores at 6 dpf obtained from zebrafish developmentally exposed to $0.04–80.0\mathrm{\mu }\mathrm{M}$ GenX Free Acid diluted in DI water daily, from $0–5$ dpf. Significance was determined by one-way ANOVA with a Tukey’s multiple comparison test ($p<0.05$). $n=6$ larvae per concentration tested. For the DNT assay, zebrafish were exposed to $4.4–80.0\mathrm{\mu }\mathrm{M}$ GenX Free Acid daily, from $0–5$ dpf. At 6 dpf, locomotor activity was assessed. (B) Distance moved (cm) each 2-min period or (C) mean distance moved during the light 1 (L1), light 2 (L2), dark 1 (D1), or dark 2 (D2) 10-min periods are shown. Repeated measures ANOVA models were run separately by period (L1, L2, D1, or D2). If a significant effect of concentration was detected ($p<0.0125$), within-period pairwise comparisons to control were computed using t-tests with a Dunnett adjustment for multiple comparisons (*$p<0.05$). $n=17–21$ zebrafish per concentration and 161 DI water control larvae were assessed. (D) Media concentrations and (E) internal tissue dose at 6 dpf following daily exposure to $25.1–80.0\mathrm{\mu }\mathrm{M}$ GenX Free Acid. $n=3$ media replicates and $n=4$ biological replicates each comprising 10 pooled larvae. Significance was determined by a Welch’s ANOVA followed by a Dunnett T3 test ($p<0.05$). Additional one-sample Student’s t-tests were performed ($p<0.05$). Note: ANOVA, analysis of variance; D, dark phase; DevTox, developmental toxicity; DI, deionized; Dil, dilution; DNT, developmental neurotoxicity; dpf, days post fertilization; GenX Free Acid, perfluoro-2-propoxypropanoic acid; L, Light period.