The GFP–G3BP1 reporter responds to oxidative and ER stresses in embryonic zebrafish. (A) Single-layer image showing GFP–G3BP1 expression in 1 dpf zebrafish under basal conditions. This region is examined in more detail in panels B and C. (B) Induction of stress granules in the retina, but not in the brain, of 1 dpf GFP–G3BP1 knock-in zebrafish after 30 mM sodium arsenite (SA) exposure in the medium for the indicated amount of time. (C) Induction of stress granules in epidermal cells in 1 dpf zebrafish exposed to 20 mM dithiothreitol (DTT) stress for indicated amount of time. (D) Induction of stress granules in the epidermal cells in 1 dpf zebrafish exposed to 10 mg/ml puromycin (PM) stress for 5 h. (E) Stress granule formation in midbrain was suppressed by treatment with 10 mg/ml cycloheximide (CHX). Enlarged images of the yellow square areas in the midbrain region are shown in the lower panels. Images show representative results from 2–3 independent experiments each with n=3–4 zebrafish examined at each condition. Scale bars: 20 µm.
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