At Montgomery, it intersects the southern terminus of I-85 and crosses the Alabama River north of the city. The Hyundai Corporation's automotive plant in Montgomery is located just off I-65. It can be accessed using the Pintlala-Hope Hull exit (number 164).
At Chilton County, I-65 enters the Birmingham, Alabama, metropolitan area. Approximately halfway between Montgomery and Birmingham, it passes Clanton (Chilton County seat), where the water tower, visible from the road, is shaped and painted to resemble a huge peach. Between mile markers 212 and 219, I-65 was designated "War on Terror Memorial Highway" in 2014.
From mile marker 242 to 290 I-65 carries at least 6 lanes of traffic. I-65 intersects I-459 in Hoover, then passes through the cities of Vestavia Hills and Homewood, which often generate heavy traffic. As the interstate passes by downtown Birmingham, south-bound travelers have a view of the Vulcan statue atop Red Mountain. At the north edge of downtown, I-65 intersects I-20/I-59 with a cross-over interchange, often called "Malfunction Junction".