Phospho h3 ph 3

The AB wild type (WT), and cox17^−/− [34 (link), 35 (link)] and atp7a^−/− (prepared in our lab, and the transcriptional profiles of the mutated embryos were organized in another manuscript) adult zebrafish were cultured in a circulating filtration system (28 ± 0.5 °C, 14,10 h light: dark). In this study, cox17^−/− and atp7a^−/− were used to test their integral function in copper induced embryonic retinal defects. Natural eggs were obtained and maintained at 28.5 °C in an incubator. The ages of the embryos and larvae were expressed by hours post-fertilization (hpf) or days post-fertilization (dpf). The following reagents and antibodies were used in the study: copper sulfate, copper nano-powders and PBA (Sigma-Aldrich, USA); NAC, reduced GSH, and RIPA lysis (Beyotime, China), and TRIzol (Life Technology Co, USA). Antibodies: Zpr-1, Zpr-3, red opsin, blue opsin, and green opsin (Abgent, USA); PDI, p­eIF-2α, and XBP1-s (Huaan Biotechnology, China), phospho-H3 (PH-3) (Cell Signaling Technology, USA).

The AB wild-type (WT) adult zebrafish were maintained in a circulation filtration system (28 ± 0.5 °C, pH 7.0–8.0, conductivity of 500–800 µS/cm) with a light:dark ratio of 14:10 h and fed with hatched fairy shrimp three times per day. Embryos were obtained by natural spawning and cultured at 28.5 °C in an incubator. The embryonic and larval ages were expressed as hours post-fertilization (hpf) or days post-fertilization (dpf). The reagents and antibodies used in the study were as follows: L-selenomethionine (Sigma-Aldrich, St. Louis, MO, USA), TUNEL Bright-Red Apoptosis Detection Kit (Vazyme, Nanjing, China), DAPI, FerroOrange (Sigma-Aldrich, St. Louis, MO, USA), NAC, Reduced GSH, Fer-1 and CDDP (MCE, Monmouth Junction, NJ, USA). Antibodies used were phospho-H3 (PH-3) (Cell Signaling Technology, Danvers, MA, USA), Caspase3 and FITC Goat Anti-Rabbit IgG (ABclonal, Wuhan, China).