Lo Truglio wrote and acted in various skits for The State, and animated segments for the show. After The State ended in 1995, he made a number of guest appearances throughout the late 1990s on shows such as Viva Variety, Upright Citizens Brigade, Law & Order and Third Watch.
In 2001, Lo Truglio appeared in fellow State alumnus David Wain's comedy film Wet Hot American Summer, where he played a camp counselor. He made cameo appearances on Wain/Showalter/Black's Stella shorts.
He appeared in the 2005 Comedy Central show Stella and had a cameo in Showalter's film The Baxter. He made cameos on Reno 911!, as well as in the 2007 movie, Reno 911!: Miami. He has provided his voice for several video games, including Grand Theft Auto: San Andreas and The Warriors. In 2005, he provided the voice of Vincenzo 'Lucky' Cilli in Grand Theft Auto: Liberty City Stories. In 2006 he had a supporting role in Artie Lange's Beer League.
He portrayed "Francis the Driver" in the Judd Apatow hit comedy Superbad and had supporting roles in films such as Pineapple Express, Paul, Role Models, Wanderlust, and I Love You, Man. In July 2008, Lo Truglio starred with Bill Hader and Jason Sudeikis in the web series The Line on Crackle. He has appeared on the comedy podcasts Comedy Bang Bang, Never Not Funny, and Superego.
Lo Truglio appeared on the Starz comedy Party Down and also had a recurring role on the short-lived 2010 Fox sitcom Sons of Tucson. In 2011, he co-starred in the short-lived NBC comedy series Free Agents starring Hank Azaria. Lo Truglio provided the voice of Freddy in the American Dad! episode "Stan's Best Friend." In 2013, Lo Truglio began co-starring opposite Andy Samberg and Andre Braugher on the Fox comedy series Brooklyn Nine-Nine.
