Reactive oxygen species (ROS) have an important pathogenic role in the development of many diseases, including kidney disease. Major ROS generators in the glomerulus of the kidney are the p47^phox-containing NAPDH oxidases NOX1 and NOX2. The cytosolic p47^phox subunit is a key regulator of the assembly and function of NOX1 and NOX2 and its expression and phosphorylation are upregulated in the course of renal injury, and have been shown to exacerbate diabetic nephropathy. However, its role in nondiabetic-mediated glomerular injury is unclear. To address this, we subjected p47^phox-null mice to either adriamycin-mediated or partial renal ablation–mediated glomerular injury. Deletion of p47^phox protected the mice from albuminuria and glomerulosclerosis in both injury models. Integrin α1–null mice develop more severe glomerulosclerosis compared with wild-type mice in response to glomerular injury mainly due to increased production of ROS. Interestingly, the protective effects of p47^phox knockout were more profound in p47^phox/integrin α1 double knockout mice. In vitro analysis of primary mesangial cells showed that deletion of p47^phox led to reduced basal levels of superoxide and collagen IV production. Thus, p47^phox-dependent NADPH oxidases are a major glomerular source of ROS, contribute to kidney injury, and are potential targets for antioxidant therapy in fibrotic disease.