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Fig. 3
Hypoxia induces ferroptosis in cerebrovascular endothelial cells. (A) The mRNA expression levels of ferroptosis-related genes in the head of zebrafish were evaluated by RT-qPCR. Mean ± SD, Student's t-test, *p < .05, **p < .01, ***p < .001. N = 80 fish per group. (B) Measurement of iron content in zebrafish head. Mean ± SD, Student's t-test, normoxia vs. hypoxia (Total Fe), **p = .0047; normoxia vs. hypoxia (Fe2+), *p = .011; normoxia vs. hypoxia (Fe3+), *p = .039. N = 150 fish per group. (C) Immunoblotting analysis of GPX4 and xCT in the head of zebrafish. Mean ± SD, Student's t-test, normoxia vs. hypoxia (GPX4), **p = .002; normoxia vs. hypoxia (xCT), *p = .034. N = 100 fish per group. (D) The mRNA expression levels of ferroptosis-related genes in bEnd.3 cells were calculated by RT-qPCR. Mean ± SD, Student's t test, *p < .05, **p < .01, ***p < .001. (E) Protein expression level of GPX4 and xCT in bEnd.3 cells was analyzed by immunoblotting analysis. Mean ± SD, Student's t test, normoxia vs. hypoxia (GPX4), ***p < .001; normoxia vs. hypoxia (xCT), *p = .014. (F) Immunofluorescence staining and quantification of GPX4. Mean ± SD, Student's t-test, **p = .0034. Scale bar: 10 μm.
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