Im Herbst 2005 übernahm Ackles eine der beiden Hauptrollen in der Mysteryserie "Supernatural" und spielt seitdem an der Seite von Jared Padalecki die Rolle des "Dean Winchester". ###>He also had a lead role in the 2005 film "Devour" in which Ackles' father, actor Alan Ackles, also had a role playing the father of Ackles' character, Jake Gray.