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Fig. 5 Hsp70 activation is critical for Cd-induced ferroptosis in vascular endothelial cells of zebrafish embryos. (A) Zebrafish embryos were pretreated with Ver (2.5 μM) and co-treated with 100 μM CdCl2. fli1a:nGFP+ ECs in the tail were observed. The scale bars indicate 500 µm and 100 µm (enlarged). The number of ECs (B) and relative area of CVP (C) were counted. The contents of LPO and MDA (D) per mg weight were examined, and the fold changes in GSH, GSSG (E) and GSH/GSSG (F) was exhibited. (G) Relative quantification of Gpx4 in Cd31+ ECs was performed. (H) Immunofluorescence illustrates CD31 and Gpx4 localization near the vessel. Scale bar = 10 µm. (n = 4). The values are presented as the mean ± SEM in histograms or as median (line), while whiskers mean min to max levels in boxplots. *P < 0.05, **P < 0.01 and ***P < 0.001 (one-way ANOVA with uncorrected Dunn’s post-test for differences between all treated groups).