Figure 5.

Release of exosomes containing⁵⁵Fe-Tf or⁵⁵Fe-His-FTH1 is independent of hepcidin regulation in BBBECs. ECs were grown on 0.4 μm pore size filters. ⁵⁵Fe-Tf (1 mg/ml) or ⁵⁵Fe-FTH1 (100 μg/ml) were added to the apical chamber and 1 μmol/L hepcidin was added to the basal chamber concurrently. Exosomes were isolated from the basal chamber media, and ⁵⁵Fe activity was measured in exosomes and supernatant fractions of control and hepcidin treatment. A and B, illustrate that hepcidin has no effect on the exosomes release of ⁵⁵Fe-Tf or ⁵⁵Fe-HisFTH1 respectively. ns nonsignificant; C and D, demonstrate that hepcidin significantly reduces supernatant ⁵⁵Fe transported from the ⁵⁵Fe-Tf or ⁵⁵Fe-HisFTH1 respectively. ⁵⁵Fe activity was measured in DPM/ml. E, illustrates immunoblot analysis of FPN1 expression in control and hepcidin treatment. The experiment was performed in three independent replicates, data are shown as the mean ± S.D. Unpaired t test. ∗p < 0.05. All experiments were performed with three technical replicates in each biological replicate. BBBECs, blood–brain barrier endothelial cells; ECs, endothelial cells; FPN1, ferroportin1; FTH1, H-ferritin; Tf, transferrin.