Figure 6

(A) Confocal image analysis of acridine orange–stained (AO-stained) and TUNEL-stained psmb1^hi2939 (50 hpf, 72 hpf, 96 hpf) mutants reveals increased apoptosis in the brain (top and middle rows, blue and white arrows) and spinal cord (bottom row, pink arrow). psmb1^–/– have increased labeling in the pharyngeal arch area (middle row, green arrow). (B and C) ImageJ (NIH) quantification of fluorescent punctae in the brain and spinal cord of mutants following AO staining (****P ≤ 0.0001, unpaired 2-tailed t test; for +/+ vs. +/–, P > 0.05). For columns left to right, n = 10, 5, 6, 2, 8, 5 (B) and n = 9, 5, 8, 2, 9, 7 (C). (D–G) Confocal imaging and quantification of AO-stained larvae (75 hpf). Exposure to BTZ (2 μM, 12–75 hpf) alone resulted in significantly increased staining in the brain. Cotreatment with EtOH and BTZ increased the number of positive dots in the brain, spinal cord, and pharyngeal arch area relative to EtOH and BTZ alone. *P < 0.05, **P < 0.01, ***P < 0.001, ****P ≤ 0.0001, 2-way ANOVA with Tukey’s multiple-comparison test. From left, column sample n = 11, 9, 13, 9 (E); n = 12, 13, 12, 11 (F); and n = 12, 10, 13, 6 (G). Scale bars: 100 μm. Data represent mean ± SD.