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Fig. 5 Effects of valproic acid (VPA) on WT-AB mitochondrial respiration at 120 h post-fertilisation (hpf) and on reactive oxygen species (ROS) accumulation at 30 hpf (n = 12 per group). (A) Mitochondrial respiratory analysis showed a decreased oxygen consumption rate (OCR) in 50 μM VPA-treated larvae compared with controls and 25 μM VPA-treated larvae. (B–D) ATP production and both maximal and basal respiration rates were reduced in 50 μM VPA-treated larvae compared with controls and 25 μM VPA-treated larvae (* p ≤ 0.05, ** p ≤ 0.01, t-test with Welch correction). Data are represented as mean ± SEM. (E) Representative fluorescence images of ROS production in controls and VPA-treated larvae, with red dotted lines indicating the region of interest. (F) Quantitative analysis of ROS production showed a significant increase in 50 μM VPA-treated larvae compared with controls and 25 μM VPA-treated larvae (**** p ≤ 0.0001, t-test with Welch correction). Data are represented as individual values (lines indicate means ± SEM).