Shrek: When Shrek and Donkey embark on their quest, Donkey starts singing "On the Road Again" before Shrek tells him to stop. After some begging, he lets him hum instead.
The Simpsons: In "Treehouse of Horror XXVIII", Homer sings a reworded version of "On the Road Again" when he's left home alone while the rest of the family goes on vacation.