Along with Squidward, Sandy, and Gary, Mrs. Puff is one of the four friends who appear at two pivotal points in the film. They are all shown in succession when SpongeBob is about to give up, and seeing his friends enslaved gives SpongeBob a dose of reality. Later, when SpongeBob frees the town, each of the four gets a triumphant scene of themselves being saved.
At the Krusty Krab, she plays a drum sting to punctuate Squidward's joke. In a deleted scene that can be watched on the Blu-ray version, Mrs. Puff is a masked boat-driver who hunts down SpongeBob and Plankton in a long and action-packed car chase.