The first appearance of the Flying Fortress in film was a Y1B-17 in the 1938 Metro-Goldwyn-Mayer (MGM) production Test Pilot, although for the crash scene a Douglas DC-2 was modified to stand in for the bomber, the Army Air Corps being unwilling to risk even having a fire lit next to the scarce type for filming.

B-17Bs of the 132nd Bomb Squadron, 9th Bomb Group, March Field, California ("Land of the Flying Fortress") were featured in the 1941 Paramount Pictures film I Wanted Wings, based on the novel of the same title by 1st Lt. Beirne Lay, Jr.

The self-explanatory 1942 Warner Bros. film Flying Fortress showed Royal Air Force Fortress Is, singly, and in formation.

B-17s appear in the 1943 RKO picture Bombardier, at Kirtland Field, New Mexico. The film follows the training of six bombardier candidates, seen through the differences between the two USAAF pilots in charge of their training over the efficacy of precision bombing.

The 1943 Warner Bros. film Air Force, directed by Howard Hawks, used at least nine B-17B, C and D model Flying Fortresses to depict the early year of World War II, including the attack on Pearl Harbor. The story revolves around an actual incident that occurred on December 7, 1941. A bomber aircrew, flying an unarmed Boeing B-17 Flying Fortress named the Mary-Ann, is ferrying the heavy bomber across the Pacific to the United States Army Air Corps base at Hickam Field, when the bomber flies right into the middle of the Japanese air attack on Pearl Harbor and the beginning of World War II.

 On April 13, 1944, Paramount Pictures released The Memphis Belle: A Story of a Flying Fortress in theaters. Directed by Hollywood legend William Wyler while he was serving in the First Motion Picture Unit of the U.S. Army Air Forces, the film tells the story of the crew of our favorite home town B-17,The Memphis Belle. 

In William Wyler's 1946 film The Best Years of Our Lives, B-17s are prominently featured. The primary male characters hitch a cross country ride in a B-17E Flying Fortress early in the story, and at the conclusion the scrapyard at Chino, California is shown full of disposal B-17s and YB-40 gunship versions of the B-17.

B-17s also figured prominently in the Oscar-winning 1949 film Twelve O'Clock High starring Gregory Peck. The film focuses on aviation leadership and the human toll in the USAAF strategy of daylight precision bombing.[102] The U.S. Air Force cooperated in the production of the film, lending aircraft to the producers and allowing filming at Eglin Air Force Base and atOzark Army Air Field.[103] The film featured an actual crash landing of a B-17, piloted by veteran stunt pilot Paul Mantz.

The other post-war (1948) film about early 8th Air Force bomber operations, MGM's Command Decision, with Clark Gableand Walter Pidgeon, relied primarily on combat footage of Flying Fortresses, although at least one B-17F and one B-17G were utilized for ground filming in California.

One ex-USAAF B-17 Flying Fortress and two ex-U.S. Navy PB-1W Flying Fortresses were retrieved from a boneyard, restored, and flown across the Atlantic Ocean for the making of the 1962 Columbia Pictures film The War Lover, based on a John Hersey novel of the same title.

Two DB-17P former drone-controllers and one B-17F were featured in the 1969 film The Thousand Plane Raid. One of the DB-17Ps briefly appeared in the bio-pic MacArthurin 1977, still wearing the same markings and paint it had for The Thousand Plane Raid.

The B-17 Flying Fortress was the subject of the 1990 Warner Bros. film Memphis Belle. During filming, one of the five vintage B-17s was destroyed in an accidental crash and a second was damaged when an engine cowling detached in flight, tearing a chunk out of the aircraft's tail (and narrowly missing a nearby P-51). There were no injuries in either incident.

Fortress is a 2012 action drama war film directed by Michael R. Phillips and released by Bayou Pictures. The film follows the crew of the Boeing B-17 Flying Fortress bomber Lucky Lass as they fly in the campaign against Italy during World War II.