Eeyore also appears in the Winnie-the-Pooh cartoons which were popularized by Disney studios. In the Disney's cartoon, he was originally voiced by Ralph Wright (who was also a major writer for Disney); his current voice actor is Peter Cullen. In Kingdom Hearts II, he is voiced by Gregg Berger.

Eeyore is often used as a beast of burden in the cartoons, most notably when he was ridden by Rabbit in his search for train "borrowers" in The Tigger Without A Name and The Pooh With A Name. Nearly all of Eeyore's houses in the cartoons have fallen down, been knocked down, or been bounced down. Eeyore is not good at rebuilding the houses; butterflies often knock them down just by landing on them. Yet he soldiers on and rebuilds them time after time. Despite his depressive nature, Eeyore is capable of great compassion, which is shown when he grows a plant that Rabbit, a master gardener, was unable to grow, just by showing the plant a little love.

Eeyore has also been featured in a number of movies: Piglet's Big Movie, The Tigger Movie and Pooh's Heffalump Movie. He appears at the Walt Disney Parks and Resorts for meet and greets. His catchphrases are "Thanks for noticin' me" and "Ohhh-kayyy."

In merchandise by The Walt Disney Company, Eeyore sometimes has an uncharacteristic smile. Also, he is somewhat less caustic and sarcastic in the Disney version than in A. A. Milne's original stories. His tail was not always fixed to him by a drawing-pin, although Disney has chosen this as part of his permanent image. Eeyore lost his tail in one of Milne's stories. Owl found it and used it as a bell pull for his door, before Winnie-the-Pooh found it for Eeyore. Christopher Robin then pinned it back on. According to Winnie the Pooh and the Honey Tree, this was possible because Eeyore is full of sawdust. Also, when Eeyore appears in animation he is colored his natural grey, though he is colored blue with a pink muzzle in merchandising.

The difference between Milne's and Disney's portrayals of Eeyore may be due to a difference between British and American culture. The original Eeyore is very British, embodying as he does a mixture of pessimism, stoicism, sarcasm and cynicism, all qualities common to the stereotypical British character. Moreover he expresses these attitudes in dry, deadpan humour, again typically British. In her book Watching the English, author Kate Fox lists "Eeyorishness" as a fundamental English characteristic. This characteristic can be found in the Disney version of Rabbit.