Fig. 4

Nitric oxide inactivates SOD to increase ROS in AA TN breast cancer cells. a-c) Cells treated with DETA-NONOate (0.5 mM and 1mM) for 48h was subjected to immunoblot blot analysis for NOX4, Mn-SOD and Cu/Zn SOD where β-actin was used as control. d) Control and DETA NONOate (1mM) treated cells were subjected to cell fractionation using kit to separate cytoplasmic and mitochondrial fraction as per manufacturer’s instructions. These fractions were further subjected to immunoblot blot analysis for NOX4, Mn-SOD, and Cu/Zn SOD. COX4 was used as positive control for purity of mitochondrial fraction. e, f, g) cells treated with DETA-NONOate (0.5 mM and 1mM) for various time points (3-36h) were subjected to SOD activity assay using kit as per manufacturer’s instructions. Data are represented in Mean ± SD with significant values presented as *p < 0.01 and **p < 0.001