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Fig. 2. Analyzing vestibular and auditory behavior of shox2 mutant larvae. (A) Representative images of shox2+/+ and shox2Δ/Δ 5 dpf zebrafish larvae. The arrow points to the swim bladder (SB). (B) Percent of embryos with inflated swim bladder at different ages in shox2+/+ (4 dpf: 72.4±13.6%, 5 dpf: 87.5±9.2%, 6 dpf: 92.6±4.4%, 7 dpf: 95.0±5.1%), and shox2Δ/Δ (4 dpf: 19.1±8.4%, 5 dpf: 23.4±6.5%, 6 dpf: 21.9±7.7%, 7 dpf: 24.3±9.9%) larvae. (C) Percent of shox2+/+ (82.7±2.8%, n=42) and shox2Δ/Δ (70.3±4.0%, n=37) larva passing the balance test (BT), embryos from different ages were combined. (D) Average distance traveled by shox2+/+ (4 dpf: 1.4±0.3 cm, 5 dpf: 3.4±0.6 cm, 6 dpf: 2.8±0.4 cm, 7 dpf: 3.4±0.5 cm) or shox2Δ/Δ (4 dpf: 2.8±0.7 cm, 5 dpf: 7.2±2.0 cm, 6 dpf: 5.6±1.0 cm, 7 dpf: 4.9±1.4 cm) larva after mechanical stimulation. (E) Percent of embryos responding to a series of tone pip in shox2+/+ (n=15) and shox2Δ/Δ (n=22) larvae. Tone bursts were presented at 0.1, 0.2, 0.3, 0.5 and 1 kHz and movements evoked from acoustic stimuli were recorded. Three independent trials were accomplished for each frequency and showed no significant (ns) differences. One-way ANOVA and Sidak multiple comparisons tests were used for statistical analysis. Values reported as mean±s.e.m. P≤0.05, **P≤0.01, ***P≤ 0.001; ****P≤ 0.0001. Values reported as mean±s.e.m.