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- Reese Witherspoon as Ginormica
- Seth Rogen as B.O.B.
- Hugh Laurie as Dr. Cockroach, PhD
- Will Arnett as The Missing Link
- Conrad Vernon as Insectosaurus
- Rainn Wilson as Gallaxhar
- Amy Poehler as Gallaxhar's Computer
- Kiefer Sutherland as General Warren R. Monger
- Stephen Colbert as President Hathaway
- Paul Rudd as Derek Dietl
- Jeffrey Tambor as Carl Murphy
- Julie White as Wendy Murphy
- Renee Zellweger as Katie
- John Krasinski as Cuthbert
- Ed Helms as News Reporter