Fig. 7.

Neither ATf iron-binding capacity nor a blockade of Tf extravasation explain protection by ATf. (A) Representative U-PAGE/WB images of the human Tf forms present in MCAO rat serum samples (R1 to R6) obtained right after the administration of hATf (AA) and 24 h later (A24). (B) Human HTf was administered i.v. to rats and TSAT of human Tf was determined in rat blood at different time-points. Human HTf retains most of its iron within the first hour after being administered, but loses most of the iron 24 h later (n = 4–5 per group; repeated measures one-way ANOVA: p = 0.0002, and SNK test, *p < 0.05, ***p < 0.001). (C) Quantification of NIR fluorescence in brains of rats exposed to tMCAO and administered NIR-hHTf* (0.8 mg/Kg) plus either Veh (green) or hATf (300 mg/Kg) (blue) at reperfusion onset. One hour later, brains were obtained and NIR fluorescence measured in the contralateral (Co) and the ipsilateral (Ip) hemispheres (n = 4–5 per group; paired t-test: *p = 0.0239 and **p = 0.0050 vs respective Co). (D) After NIR imaging, brains in (C) were processed and representative WB shows bands corresponding to exogenous hATf and actin, in Co and Ip hemispheres of Veh and hATf-treated rats 1 h after reperfusion. (E) Quantification of hATf levels in WB of brain hemispheres (n = 4–5 per group; paired t-test: *p = 0.0500).