Effects of parental treatment on zebrafish larvae cellular stress protein expression. Control and post-exposure to combined elevated temperature and hypoxia representative western blots and relative protein expression for (A) HSP70, (B) HSP90 and (C) HSP47 in 5 days post-fertilization (dpf) larvae derived from adult zebrafish exposed to either control (offspring control) or combined exposure conditions (offspring combined exposure) for 14 days. Western blot bands and protein expression was normalized and expressed as stated in Fig. 5. All protein expression was further normalized to control larvae derived from control parents. Statistical differences between values were determined by two-way ANOVA followed by a Holm–Šidák post hoc test (A: parental exposure: P=0.948; larval exposure: P<0.001; parental exposure×larval exposure: P=0.815), or two-tailed t-test (C: within control: P=0.941) or Mann–Whitney rank sum test (B: within control: P=0.394, within combined exposure: P=0.937; C: within combined exposure: P=0.002) across exposure at 5 dpf, followed by two-tailed t-tests (B: offspring control: P=0.987, C: offspring Control: P<0.001, offspring combined exposure: P=0.015) or Mann–Whitney rank sum tests (B: offspring combined exposure: P=0.394) between parental treatments. Differences within treatments across exposure at 5 dpf are indicated by different letters. Differences between treatments for a given exposure at 5 dpf are indicated by an asterisk. Data are means±s.e.m. in A, and medians and interquartile ranges in B and C (all n=6, except HSP70 for combined exposure larvae from control parents, n=5).
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