Deletion of the Ciao3 Gene Leads to Embryonic Death and Vascular Development Disorder in Mice. A) The embryo morphology of the Ciao3 heterozygous offspring at 8.5, 10.5, 12.5 and 13.5 days; Ciao3 knockout embryos still existed at 12.5 days but were completely absorbed at 13.5 days and later. B) H&E staining morphology of wild type with 8.5‐day and 12.5‐day mouse embryos and Ciao3−/− mice embryos with 10.5‐day and 12.5‐day, the development of Ciao3 knock‐out embryos was significantly slower than that of wild‐type mouse embryos and the development of the vascular system was blocked. C) The endothelial marker CD31 was detected by immunofluorescence staining in 12.5‐day mouse embryos. D) The endothelial progenitor cell marker CD34 was detected by immunofluorescence staining in 12.5‐day mouse embryos. E,F) The sections of 12.5‐day‐old mouse embryos were detected by 4‐HNE (E) and BODIPY (F) probes; the positive rate of 4‐HNE and BODIPY in Ciao3−/− mouse embryo was significantly higher than in wild‐type mouse embryo. H) γ‐H2AX detection of 12.5‐day mouse embryos showed that the DNA damage of Ciao3−/− mouse embryos was significantly enhanced compared with wild‐type embryos. G‐I) Western blotting showed that GPX4, xCT and FTL were significantly down‐regulated in Ciao3 knockout mouse embryos; the expression of TFR1 and IRP1 was significantly up‐regulated. Data were analyzed by an unpaired Student's t‐test; *p < 0.05; **p < 0.01; ***p < 0.001.
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