Luke Air Force Base serves as a primary training site for both US and international pilots transitioning from older aircraft like the F-16 Fighting Falcon to the newer F-35. It houses five F-35 squadrons dedicated to pilot training.

Nellis Air Force Base is recognized for hosting air combat exercises such as Red Flag. It supports units like the 422nd Test and Evaluation Squadron operating F-35s. Edwards Air Force Base also plays a crucial role as it hosts test units including NASA's Armstrong Flight Research Center.

Eglin Air Force Base focuses on joint graduate flying and maintenance training through its 33rd Fighter Wing "Nomads," which includes units like the 58th Fighter Squadron "Mighty Gorillas."

In addition to these locations, Marine Corps and Navy bases are also equipped with F-35 variants suitable for their operations on amphibious assault ships and aircraft carriers. The Marines operate from bases such as Marine Corps Air Station Beaufort in South Carolina and Marine Corps Air Station Yuma in Arizona. The Navy operates from locations including Naval Air Station Patuxent River in Maryland and Naval Air Station Lemoore in California.

The UK's Royal Air Force Lakenheath serves exclusively US forces with four fighter squadrons—two of which fly the F-35A while two continue with the F-15E Strike Eagle until their transition to the new jets.

Despite challenges such as unidentified drones flying over military bases, no direct threats have been reported so far. As part of global defense infrastructure enhancement efforts, many allied nations are integrating their own fleets of this advanced fifth-generation fighter jet into their air forces.