Fig. 4.

Ascorbate, dehydroascorbic acid (DHA) and bromoDHA transport into mouse RBCs. A. RBC ascorbate was measured in RBCs that were incubated with ascorbate (100 μM) for 0–10 min at 0 °C or 37 °C. B. Freshly prepared DHA (100 μM) was added for 0–10 min to RBCs at 37 °C with and without cytochalasin B pre-treatment and to RBCs at 0 °C with and without cytochalasin B pre-treatment. C. Freshly prepared bromoDHA (100 μM) was added for 0–10 min to RBCs at 37 °C and 0 °C; controls were freshly prepared DHA (100 μM) added for 0–10 min to RBCs at 37 °C and 0 °C. D. BromoDHA (100 μM) was kept for 0–10 min at 37 °C, and then reduced to bromoAA for measurement. E. BromoAA and ascorbate binding to unsealed ghosts of wildtype mouse RBCs. Unsealed ghosts prepared as in Methods from 200 μL of wildtype mouse RBC were incubated with 100 μM bromoAA or AA for 20 min at 37 °C. Ghost pellets were obtained by centrifugation, and ascorbate and bromoAA were measured by HPLC (n = 3).