Fig. 5. Synergistic toxicity of iron and inflammation is prevented by αTocopherol treatment.

a Primary HUVECs were treated with solvent (gray gircles), 100 μM ferric ammonium citrate (FAC) (light blue circles), or 100 μM FAC with vitamin E analog Trolox (300 μM, white circles) for 24 h prior to being stimulated with TNFα (50 ng/ml) for 16 h. Western blot (left) and quantitation (right) of cleaved caspase-3 normalized to β-actin (representative image of n = 3 independent experiments). b–i Hepcidin KO dams were treated with subcutaneous solvent (dark blue circles) or αTocopherol (αToc: vitamin E, white circles) 14 and 2 h prior to LPS treatment on E15.5 for 24 h: c embryo gross morphology and incidence of resorbing embryos; d maternal liver Il6 expression relative to Hprt from n = 6 dams; e–g placental Il1b, Cxcl2, and Il6 expression relative to Rpl4 from n = 10 placentas. h Western blot (left) and quantitation (right) of cleaved caspase-3 in whole placentas normalized to β-actin. n = 6 placentas/group. i Immunohistochemistry for cleaved caspase-3 (brown) and CD31 (red) in paraffin-embedded placenta, embryo lung and liver (representative image of n = 3 sections/group). Scale bar = 50 μm. a, c–h Error bars represent mean ± s.e.m. e–i Embryo and placentas were randomly selected for analysis. Statistical differences were determined by two-tailed Mann–Whitney U, two-tailed Student’s t-test (denoted by *) or one-way ANOVA followed by Holm-Sidak method for multiple comparisons (denoted by #). P-values are indicated in each figure panel. Source data are provided as a Source data file.