- The Other Darrin: Several characters switched voice actors between the cartoon and the game such as Invisible Woman and Storm going from Tara Strong and Cree Summer to Grey DeLisle. In addition, some characters switched voice actors during the game's life such as Spider-Man going from Mikey Kelly to Yuri Lowenthal with the latter going to voice the web-head several more times. Another example is Phil Coulson, who was voiced by Tom Kenny rather than Clark Gregg, thus making him the only other actor to have ever played Coulson.
- Role Reprise: In addition to most of the show's voice cast.
- Grey DeLisle reprises her role from the bonus levels of Fantastic Four (2005)'s tie-in game as the Invisible Woman.
- David Sobolov, Nika Futterman, and James C. Mathis III reprises their roles from the Marvel Universe shows as Drax the Destroyer, Gamora, and Ronan the Accuser.
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