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Heat shock-induced cell death coincides with decreased level of p-eIF2α and reduced number of stress granules in fish larvae. (A) Representative western blots showing the expression of phosphorylated eIF2α (p-eIF2α), total eIF2α (t-eIF2α) and actin after 10 min heat shock in the brain of 1, 3 and 11 dpf GFP–G3BP1 knock-in zebrafish. (B–D) Quantification of the expression of t-eIF2α relative to actin (B), p-eIF2α relative to actin (C) and p-eIF2α relative to p-eIF2α (D). Values represent mean±s.e.m. Data from three independent experiments, with n=15–20 zebrafish brains pooled for protein analysis for each condition. *P≤0.05, **P≤0.01 by unpaired Student's t-test. (E) Representative images showing stress granule formation and cell death (revealed by TUNEL staining, red) in the epidermis of 1, 3 and 11 dpf GFP–G3BP1 knock-in zebrafish exposed to 42°C for 20 min. n=4–5 zebrafish at each age with similar results. Scale bar: 10 µm.
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