- Audrey Hepburn was first offered the role of Anne Frank. She refused it for two reasons: she had decided to accept the role in Green Mansions (1959) and, more importantly, she had lived in occupied Holland during the war and had seen the Nazis carry out street executions and watched as they herded Jews onto boxcars to carry them to concentration camps. She knew that making the film would bring back memories that were far too painful for her.
- Stevens had trouble getting suitable emotion from the actors when the American plane passes by. So, a record player was hidden in the sound stage and was set to play "The Star Spangled Banner" to evoke an emotional reaction from the American actors.
- All annex scenes were shot on the largest sound stage at 20th Century Fox in Hollywood. The real annex would have been too small to accommodate a film crew.
- Shelley Winters had to gain 25 pounds for the role of Mrs. Van Daan. She then had to lose 15 of it as the movie progressed.
- Shelley Winters donated the best supporting actress Academy Award she won for the role of Mrs. Van Daan to the Anne Frank museum in Amsterdam.
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