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Elimination of Young Erythrocytes from Blood Circulation and Altered Erythropoietic Patterns during Paraquat Induced Anemic Phase in Mice

Figure 2

Effect of paraquat on erythrocyte turnover.

Double biotinylation technique was used to label erythrocytes in vivo as described in methods. Mice were administered three daily i.v. doses of 1 mg biotin-X-NHS Ester (BXN) (first biotinylation step). After 5 days rest, a single i.v. dose of 0.6 mg BXN was administered (second biotinylation step). Repeated doses of paraquat (10 mg/kg) were administrated i.p. on alternate days starting the day after the second biotinylation step. Blood samples were collected on day 0 just before the first paraquat injection, and at 7, 14, 21 and 28 day time points during the paraquat administration. Erythrocytes in the blood samples were stained with streptavidin-APC and analyzed on flow cytometer. Box-Y in panel A and B contain erythrocytes that entered the circulation between the first and the second biotinylation step in control and paraquat treated mice respectively. Erythrocytes younger than those in box-Y are in box-Z (biotinnegative population) whereas the erythrocyte population older than the population in box-Y is represented as events in box-X (biotinhigh population). Panel C shows the survival kinetics of older erythrocytes (box-X) in control and paraquat treated mice. Panel D shows the kinetics of survival of the tracked cohort of box-Y erythrocytes in control and paraquat treated mice. Panel E shows the kinetics of accumulation of younger box-Z erythrocytes in control and paraquat treated mice. Data is represented as mean ± SEM; n = 6 in control and 4–5 in treated groups. *p<0.05, **p<0.005 and ***p<0.0005.

Figure 2

doi: https://doi.org/10.1371/journal.pone.0099364.g002