Fig. 2.

Calcium deficiency abates catalase activity and increases ROS accumulation. A Catalase activity assay of AML 12 cells in calcium deficient medium for indicated durations. B AML12 cells in calcium-deficient medium for indicated durations were stained with Cell ROX. Red fluorescence represents the ROS. Scale bar represents 150 µm. C Quantification of relative red fluorescence intensity indicates ROS content. Data are expressed as means ± S.D. (n = 3, independent experiments, 30 cells were analyzed in each experiment), * p < 0.05. D AML12 cells in calcium-deficient medium for indicated durations treated with antioxidant NAC were stained with Cell ROX. Scale bar represents 150 µm. E Quantification of relative red fluorescence intensity indicates ROS content. Data are expressed as means ± S.D. (n = 3, independent experiments, 30 cells were analyzed in each experiment), * p < 0.05. F Chang Hyper-SKL cells in calcium deficient medium for indicated durations. Green fluorescence represents ROS content from peroxisomes. G Quantification of green fluorescence intensity indicates peroxisomal ROS content. Data are expressed as means ± S.D. (n = 3, independent experiments, 30 cells were analyzed in each experiment), * p < 0.05. H Chang Hyper-SKL cells in calcium deficient medium treated with antioxidant NAC for indicated durations. Green fluorescence represents the ROS content in peroxisomes. Scale bar represents 150 µm. I Quantification of green fluorescence intensity indicates peroxisomal ROS content. Data are expressed as means ± S.D. (n = 3, independent experiments, 30 cells were analyzed in each experiment), * p < 0.05