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Fox was also cast in a regular role on the ABC sitcom Hope & Faith, in which she portrayed Sydney Shanowski, replacing Nicole Paggi.

Fox appeared in seasons 2 to 3, until the show was cancelled by ABC in May 2006.

She attended a dance class at the community center there and was involved in Kingston Elementary School's chorus and the Kingston Clippers swim team. She said that the problem was not her looks, but that she had "always gotten along better with boys" and that "rubbed some people the wrong way".

Fox also said of high school that she was never popular and that "everyone hated me, and I was a total outcast, my friends were always guys, I have a very aggressive personality, and girls didn’t like me for that. In the same interview, she mentions that she hated school and has "never been a big believer in formal education" and that "the education I was getting seemed irrelevant. At 15, Fox made her acting debut in the 2001 film Holiday in the Sun, as spoiled heiress Brianna Wallace and rival of Alex Stewart (Ashley Olsen), which was released direct-to-DVD on November 20, 2001.

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Scholars Wheeler Winston Dixon and Gwendolyn Audrey Foster disagreed that every woman in Hollywood is marketed as a sex symbol, and stated that Fox's "celebrity is based on what she admits is an entirely artificial construct designed for dissemination in the Internet age, a 21st-century media personality in every sense of the word." Fox's public image came under scrutiny when, on September 11, 2009, an unsigned letter from three crew members of Transformers defended Michael Bay against accusations made by Fox about his on-set behavior, including a comparison with Hitler.

In 2007, Fox won the lead female role of Mikaela Banes in the 2007 live-action film Transformers, based on the toy and cartoon saga of the same name.

Fox played the love interest of Shia La Beouf's character Sam Witwicky.

In 2012, Fox appeared briefly in Sacha Baron Cohen's comedy The Dictator and had a featured role in Judd Apatow's comedy This Is 40.

She was the voice of Lois Lane in the film Robot Chicken DC Comics Special, an episode of the television comedy series Robot Chicken, and it aired as a one-off special during Cartoon Network's Adult Swim on September 9, 2012.