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. 2020 Jul 13;15(7):e0235491. doi: 10.1371/journal.pone.0235491

Fig 2. SIRT3 loss of function impacts initial auditory thresholds but not hearing recovery from TTS.

Fig 2

a. The time course of experiments is shown. Homozygous Sirt3-KO mice and wild-type littermates are tested for ABR and DPOAE thresholds prior to noise exposure (pre-test). At P60, they are exposed to an 8–16 octave band noise at 105 dB for 30 minutes (noise), which is sufficient to drive temporary threshold shifts in this strain [21, 23]. Their hearing thresholds are assessed again at 1 day post noise (1 DPN) and 14 days post noise (14 DPN). ABR and DPOAE are assessed at 8, 12, 16, 24, and 32 kHz for all panels. b. Mean ABR thresholds prior to noise exposure for homozygous Sirt3-KO mice (n = 14, pink) and wild-type littermates (n = 13, black). Mice were approximately P50 when tested. Homozygous Sirt3-KO mice had significantly worse overall hearing (p = 0.004 for genotype, two way ANOVA, n = 27 mice total, noted as **); however, no single frequency was significantly worse (p = 0.053, 0.39, 0.41, 0.53, 0.059 for the five frequencies respectively, two-tailed pairwise t-tests at each frequency with Bonferroni correction, n = 27 mice total). c. Mean ABR thresholds at 1 DPN for homozygous Sirt3-KO mice (n = 14, pink) and wild-type littermates (n = 13, black). No significant difference was seen between genotypes (p = 0.35 for genotype, two-way ANOVA, n = 27 mice total, noted as n.s.). d. Mean ABR thresholds at 14 DPN for homozygous Sirt3-KO mice (n = 14, pink) and wild-type littermates (n = 13, black). No significant difference was seen between genotypes (p = 0.85 for genotype, two way ANOVA, n = 27 mice total). Overall ANOVA for all three time points showed no significant difference between genotypes (p = 0.33, multi-way ANOVA, n = 27 mice total, noted as n.s.). e. Mean DPOAE thresholds prior to noise exposure for homozygous Sirt3-KO mice (n = 14, pink) and wild-type littermates (n = 13, black). No significant difference was seen between genotypes (p = 0.43 for genotype, two-way ANOVA, n = 27 mice total, noted as n.s.). f. Mean DPOAE thresholds at 1 DPN for homozygous Sirt3-KO mice (n = 14, pink) and wild-type littermates (n = 13, black). No significant difference was seen between genotypes (p = 0.50 for genotype, two-way ANOVA, n = 27 mice total, noted as n.s.). g. Mean DPOAE thresholds at 14 DPN for homozygous Sirt3-KO mice (n = 14, pink) and wild-type littermates (n = 13, black). No significant difference was seen between genotypes (p = 0.25 for genotype, two-way ANOVA, n = 27 mice total, noted as n.s.). Error bars: s.e.m.