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. 2015 Oct 3;2(11):1735–1750. doi: 10.1016/j.ebiom.2015.09.049

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This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Fig. 1 — Hypothesis origin and vitamin C concentrations in RBCs and plasma.

A. Hypothesis origins: findings from centrifugation of mouse whole blood. For ascorbate values, samples were obtained from 5 Gulo^−/− mice supplemented with ascorbate 1 g/L in drinking water for 12 weeks and 5 Gulo^−/− mice not supplemented with ascorbate for 12 weeks, plasma concentrations indicated in the panel. Gulo^−/− mice do not make ascorbate. When ascorbate was provided, mice received it via drinking water, which was changed daily.

B. Mouse RBC ascorbate as a function of plasma ascorbate. 12 wildtype (WT) mice were unsupplemented for 14 weeks (); 10 Gulo^−/− mice were unsupplemented from 0 to 14 weeks (); and 5 Gulo^−/− mice were supplemented with 1 g/L ascorbate in drinking water from 0 to 17 weeks (●). Each symbol represents a separate blood sample. R = 0.91, p < 0.01.

C. Human RBC ascorbate as a function of plasma ascorbate in 153 healthy blood donors, ascorbate measured by HPLC with coulometric electrochemical detection (Li et al., 2012). Each sample was measured in triplicate, samples without error bars indicate SD was less than symbol size. R = 0.82, p < 0.02.

D. Human mononuclear cell ascorbate (all non-open circle symbols) as a function of plasma ascorbate, ascorbate measured by HPLC. In-patient healthy subjects (6 men and 13 women) who were at steady state for vitamin C doses of 30, 60, 100, 200, 400, 1000, and 2500 mg in two divided doses daily underwent apheresis with elutriation of cell-enriched product to obtain mixed mononuclear cells, as described (Levine et al., 1996, Levine et al., 2001). Each symbol represents a different subject. For each subject, samples were obtained at 1–5 different vitamin C doses at steady state for that dose (Levine et al., 1996, Levine et al., 2001).

Inline graphic — Hypothesis origin and vitamin C concentrations in RBCs and plasma.

A. Hypothesis origins: findings from centrifugation of mouse whole blood. For ascorbate values, samples were obtained from 5 Gulo^−/− mice supplemented with ascorbate 1 g/L in drinking water for 12 weeks and 5 Gulo^−/− mice not supplemented with ascorbate for 12 weeks, plasma concentrations indicated in the panel. Gulo^−/− mice do not make ascorbate. When ascorbate was provided, mice received it via drinking water, which was changed daily.

B. Mouse RBC ascorbate as a function of plasma ascorbate. 12 wildtype (WT) mice were unsupplemented for 14 weeks (); 10 Gulo^−/− mice were unsupplemented from 0 to 14 weeks (); and 5 Gulo^−/− mice were supplemented with 1 g/L ascorbate in drinking water from 0 to 17 weeks (●). Each symbol represents a separate blood sample. R = 0.91, p < 0.01.

C. Human RBC ascorbate as a function of plasma ascorbate in 153 healthy blood donors, ascorbate measured by HPLC with coulometric electrochemical detection (Li et al., 2012). Each sample was measured in triplicate, samples without error bars indicate SD was less than symbol size. R = 0.82, p < 0.02.

D. Human mononuclear cell ascorbate (all non-open circle symbols) as a function of plasma ascorbate, ascorbate measured by HPLC. In-patient healthy subjects (6 men and 13 women) who were at steady state for vitamin C doses of 30, 60, 100, 200, 400, 1000, and 2500 mg in two divided doses daily underwent apheresis with elutriation of cell-enriched product to obtain mixed mononuclear cells, as described (Levine et al., 1996, Levine et al., 2001). Each symbol represents a different subject. For each subject, samples were obtained at 1–5 different vitamin C doses at steady state for that dose (Levine et al., 1996, Levine et al., 2001).