The route additionally co-signs with Interstate 70 and crosses the Ohio River on the Fort Henry Bridge in Wheeling, West Virginia.

In Moundsville, West Virginia, the route leaves WV 2 and departs toward Cameron, Mannington, and Fairmont.

U.S. Route 250 Truck follows Blue and Gray Expressway around the south and west sides of downtown Philippi, avoiding the covered bridge.

US 250 runs 166.74 miles (268.34 km) from the West Virginia state line near Hightown east to its eastern terminus at US 360 in Richmond.

From Staunton east to Richmond, the U.S. Highway serves as the local complement to Interstate 64 (I-64), roughly following the 18th century Three Notch'd Road through Waynesboro and Charlottesville on its way through the Shenandoah Valley, its crossing of the Blue Ridge Mountains at Rockfish Gap, and the Piedmont.

This well-planned route required only one major river crossing, the Rivanna at Charlottesville, with inns or taverns spaced about 10 miles (16 km) apart.

In the second quarter of the 19th century, it became a major gateway into Virginia's "Trans-Allegheny" region, which presented much greater challenges to transportation than did less rugged portions of the state.

As the American Civil War broke out, the route became very important and was defended by Stonewall Jackson's troops at the Battle of McDowell in 1862.