Harley Quinn is the Joker's female sidekick and one of Pooh's enemies. Her first appearance in a Pooh's Adventures film was Winnie the Pooh Goes to Atlantis: The Lost Empire where she, The Joker, The Penguin, and Two-Face worked for Commander Rourke.
- Harley and the Joker later appeared in the Pooh's Adventures of Teenage Mutant Ninja Turtles and Pooh's Adventures of Teenage Mutant Ninja Turtles 2: The Secret of The Ooze where they worked for The Shredder.
- Harley and The Joker recently appeared in Pooh's Adventures of 101 Dalmatians where they worked for Cruella De Vil along with Jafar, Maleficent, Iago, Ursula, Hades, and The Grand Duke of Owls.
- Harley and The Joker will return in Pooh's Adventures of Aladdin and The King of Thieves.