- Owen Wilson as Lightning McQueen, described by John Lasseter in the LA Times as "A hybrid between a stock car and a more curvaceous Le Mans endurance racer."
- Paul Newman as Doc Hudson, a 1951 Hudson Hornet, later revealed to be the Fabulous Hudson Hornet.
- Bonnie Hunt as Sally Carrera, a 2002 996-series Porsche 911 Carrera.
- Larry the Cable Guy as Mater, a 1951 International Harvester L-170 "boom" truck with elements of a mid-1950s Chevrolet. One-Ton Wrecker Tow Truck.
- Tony Shalhoub as Luigi, a 1959 Fiat 500. *
- Cheech Marin as Ramone, a 1959 Chevy Impala Lowrider.
- Michael Wallis as Sheriff, a 1949 Mercury Club Coupe (police package).
- George Carlin as Fillmore, a 1960 VW Bus.
- Paul Dooley as Sarge, a 1941 Willys model jeep, in the style used by the US Military.
- Jenifer Lewis as Flo, a 1957 Motorama show car.
- Guido Quaroni as Guido, a custom forklift, resembling an Isetta at the front.
- Richard Petty as Strip "The King" Weathers. The car's design was based on Richard Petty's 1970 Plymouth Superbird
- Michael Keaton as Chick Hicks, described by Pixar as a generic 1980s stock car. Strongly resembles a 1978–88 General Motors G-Body such as a Buick Regal or Chevrolet Monte Carlo.
- Katherine Helmond as Lizzie, a 1923 Ford Model T.
- John Ratzenberger as Mack, a 1985 Mack Super-Liner.
- Joe Ranft as Red, a 1960s style fire truck (most closely resembles a mid-1960s), this was Ranft's last voice role before his death in August 2005.
Radiator Springs and vicinity
- The Cozy Cone Motel's design is based on the two Wigwam Motels along Route 66, in Holbrook, Arizona and Rialto, California. These were once two out of seven built motels, with individual cabins shaped like teepees. Three Wigwam Motels remain; the third (and oldest) is in Cave City, Kentucky, far from Route 66. The recently restored Tee Pee Motel in Wharton, Texas, south of Houston, is of similar design but unrelated. The name "Cozy Cone" was inspired by the Cozy Dog Drive-In of Springfield, Illinois, which lays claim to being birthplace of the corn dog.
- The character "Fillmore", referring to the famous San Francisco music venue The Fillmore, was at one time to be named "Waldmire" after Bob Waldmire, a self-proclaimed hippie artist known to Rt. 66 fans for his detailed pen-and-ink maps and postcards of the route. Though Waldmire's family owns the Cozy Dog Drive-In, Bob, having since become a vegan, preferred not to see his name put on a character that would become a McDonald's Happy Meal toy.
- Ramone's House of Body Art is based primarily on the U-Drop Inn in Shamrock, Texas. It opened in 1936 as Tower Conoco (from its distinctive Art Deco spire) with the U-Drop Inn Cafe and a retail building attached. Many other establishments built along Route 66 in its heyday had Art Deco elements that might be reflected in the design of Ramone's.
- The yellow billboard for Lizzie's Curio Shop reading "HERE IT IS" with an image of a Model T is based on the Jack Rabbit Trading Post signage in Joseph City, Arizona.
- Sheriff is voiced by Michael Wallis, an American historian and author of Route 66: The Mother Road.