At the beginning of the film when Slinky is talking about how Woody never steered them wrong before, Mr. Potato Head takes off his mouth and pats it against his rear in a sarcastic manner. In case you don't get it, he's indicating that Slinky is ass-kissing Woody.
The censors didn't mind Woody calling Buzz "Mr. Lightbeer." However, when it was broadcasted on ABC Family recently, they dubbed it over with another line from the movie, in which he says "lightsnack".
Kari: What's the S [on his shirt] for? Syndrome: For... "Sitter"! Yeah, "Sitter". At first I was thinking of getting initials for "Baby Sitter", but then I would be walking around with a big "BS" and *starts laughing* you understand why I couldn't go with that...
The attempted suicide at the beginning of the movie.
Dash's disgust when he realizes that he had been sleeping on top of Violet.
For a while, Helen thinks Bob is having an affair. The DVD reveals that there was originally going to be a scene where Helen confronts him directly with her suspicions, but that it was cut because it felt a bit too adult. Another deleted scene from the same plot ark featured Helen having a nightmare in which a newly-fit Bob is being attended to by a harem of women.
Lightning McQueen gets towed several times by Mater. Each time the tow hook is swung under his bumper, he gets a comical expression of surprise and a squeak of pain.
Lizzy a habit of slapping "Nice Butte" bumper stickers on unsuspecting male...er, bumpers. Each time she does this, it is accompanied by the same surprised squeak.
Lizzy has a line where she tells one of the other cars to move over so that she can "get a look at that sexy hotrod" (referring to Lighting).
There was also a scene where a couple of giggling teen girl cars flash their headlights at Lightning. Especially notable when you remember what part of a woman's anatomy "headlights" is a euphemism for. A deleted scene has these two as waitresses at a seedy truck stop Mack stops at on the drive to California. It was marked as a "top-down" truck-stop, with all-convertible waitresses.
The ending with the two minivans: "Oh for the love of Chrysler, can we please ask someone for directions?!"
Mater's line "He did what in his cup?!" after Lightning mentions the Piston Cup. And that was in the trailer.
There are several moments near the beginning of the movie, when Lightning makes the appearance for his sponsors. When we first see inside the Rust-eze tent, a pair of Click and Clack-esque spokescarsnote played by the real Click and Clack from Car Talk are performing a comedy set; their routine contains — among other things — a rather obvious Double Entendre:
Van: Winter is a grand old time! Rusty Rust-eze: Of this there are no ifs or buts! Dusty Rust-eze: But remember, all that salt and grime... Rusty: Can rust your bolts and freeze your— (Lightning appears) Dusty:Hey, look! There he is!
After Lightning is introduced by the Rust-eze guys, one of the fans in the crowd exclaims, "That race was a pisser!" (No kidding — it's in the actual script.)
In another scene, Lightning bursts into Doc Hudson's office and interrupts a physical exam — getting a view of the Sheriff's exposed undercarriage in the process. The Sheriff angrily barks, "Get a good peek, city boy?" (And later, as Lightning leaves the office, calls out, "Hope you enjoyed the show!")
The Tear Jerker scene at the beginning where Carl and Ellie are making a baby's room, only for it to transition to Ellie crying in the doctor's office about not being able to have a child. We don't know if it's a miscarriage or if she's infertile, but who expected that to come from Pixar!?
Each cast member mentioned in the credits is accompanied by a scrapbook-type picture that roughly corresponds with what they're being credited for (a picture of Carl & Russell making music with drinking glasses for "Music", for example). The picture for "Production Management"? Carl & Russell surrounded by Dug's puppies!
At the end of Tokyo Mater, the villain Kabuto (an obnoxious Japanese racing car) is stripped of his modifications as a result of him losing to Mater in a drift race.
In the short film El Materdor (where Mater imagines himself a bulldozer fighter) at the end he is between "the twins," teenage girl Miatas Mia and Tia, who are fawning over him. He swings his hook over and yanks one of the girls closer by her rear bumper. Her surprised facial expression, though momentary, pretty much confirms that he's grabbed her butt.