Figure 5

(A) Larval survival at 4 dpf after treatment with 1% EtOH (12 hpf–4 dpf) or 2.5 μM BTZ. Survival significantly decreases following exposure to 1% EtOH and BTZ in combination. (B) Edema prevalence in 5 dpf larvae exposed to a EtOH, BTZ, or both in increasing concentrations. For A and B, P values determined by a 2-sided Fischer’s exact test (*P < 0.05, **P ≤ 0.01, ****P ≤ 0.0001). (C) Alcian blue staining of 0% and 1% EtOH-treated (12 hpf–4 dpf) larvae in the presence of DMSO, BTZ, and MG132. BTZ and MG132, especially in the context of 1% EtOH exposure, induced Meckel’s cartilage malformations (black arrows). (D) Quantification of the percentage of affected individuals with Meckel’s cartilage abnormalities from each condition (**P < 0.01, 2-sided Fischer’s exact test). BTZ concentration = 2.5 μM (12 hpf–4 dpf). MG132 concentration = 5 μM (12 hpf–4 dpf). Scale bars: 100 μm. For D, percentages are based on n > 30 per group. See Supplemental Tables 4 and 5 for raw data and statistics related to A and B.