Fig 4. Generation of Reactive Oxygen Species (ROS) in control and cadmium treated mice.

Mice were given cadmium chloride dissolved in drinking water (50 ppm and 1000 ppm). Erythrocytes were stained with CMH₂DCFDA and the ROS generation was determined by flow cytometry. ROS generation in control mice was taken as 100 and the ROS levels in exposed groups were estimated in relative terms Relative ROS generation in mice exposed to 50 ppm (panel A) and 1000 ppm (panel B) of cadmium chloride are shown in the above panels. Mouse erythrocytes labeled in vivo by the two step biotinylation procedure were stained with CMH₂DCFDA, Streptavidin-APC and anti-mouse CD71-PE, and the ROS generation in different subpopulations of erythrocytes was determined. The percent change in ROS generation as compared to control in reticulocytes, biotin^negative, biotin^low and biotin^high erythrocytes in control and cadmium exposed mice, after 3 weeks of cadmium exposure have been shown in panel C. Each point and each bar on the graph represents mean ± SEM of observations on 7–10 mice. *p<0.05, **p<0.01 ***p<0.005 and ****p<0.001 for comparison of the groups. Statistical analysis was done using Student t-test.