Fig. 1

In ovo BHP injection increases mortality, mitochondrial ROS accumulation and oxidative damage in embryonic liver. A Developmental alteration in chick embryo in response to in ovo injected BHP levels after 24 h. The BHP concentrations of in ovo injection were 0 (pbs), 150, 300, 600, and 1200 μmol/L BHP per kg egg weight. B Embryonic mortality induced by BHP in ovo injection in a dose dependent manner. The mortality was up to 100% observed at 1200 μmol/L BHP group. C Representative hepatic morphological changes in embryo by HE staining. Black arrows represent the swollen and damaged hepatic tubule and glomerulus (black arrows). All micrographs 200 × magnification, hematoxylin/eosin staining. D Increase of hepatic MSD induced by BHP in ovo injection. E Hepatic mtROS accumulation induced by in ovo BHP injection. F Decrease of hepatic MMP by in ovo BHP injection. G–H Aged changes of hepatic mitochondrial ROS and MMP induced by BHP in ovo injection. I Lipid peroxidation assessed by MDA production in isolated cytoplasm and mitochondria. J The mtDNA damage assessed by 8-OHdG production in isolated cytoplasm and mitochondria. Graph bars in B, D, E, G, H, I and J marked with different letters on top represent statistically significant results (P < 0.05) based on Tukey's post hoc one-way ANOVA analysis, whereas bars labelled with the same letter correspond to results that show no statistically significant differences. The results from G and H were analyzed using unpaired two-tailed Student's t-test (^∗P < 0.05). Data are mean ± SEM (n = 3 or 4)