Elimination of Young Erythrocytes from Blood Circulation and Altered Erythropoietic Patterns during Paraquat Induced Anemic Phase in Mice
Mice were treated with paraquat (10 mg/kg i.p. on alternate days) for 28 days. At different given time points, blood samples were taken and ROS levels in erythrocytes was estimated flow cytometrically after treatment with CMH2DCFDA dye as described in methods. Summarized time kinetics data from 5 mice, depicting relative levels of ROS in erythrocytes in response to paraquat treatment are shown in panel A where control MFI due to ROS generation in control erythrocytes was taken as 100 and ROS levels in paraquat treated mice depicted in relative terms [*p<0.05 and **p<0.005]. Representative histograms showing the levels of ROS in erythrocytes of different age groups from control and paraquat treated mice are shown in panels B1 to B6. Mice were DIB labeled and treated with paraquat as described in legends to Figure 2. After 14 days of the paraquat treatment, blood cells were stained with streptavidin-APC to delineate the biotinnegative, biotinl°w and biotinhigh subpopulations of erythrocytes. For ROS estimation, cells were incubated with CMH2DCFDA dye as described in methods. Fluorescence generated in biotinnegative subpopulation (younger subpoplation, panel B1, B2), biotinl°w subpopulation (intermediate age subpopulation, panel B3, B4) and biotinhigh subpopulation (older erythrocyte subpopulation, panel B5, B6) was estimated flow cytometrically.