Ito presided over the 1995 murder trial of O.J. Simpson, at which Simpson was acquitted. His decision to allow television coverage of the trial was controversial, and Ito faced criticism for seeming to enjoy the press and for allowing too many sidebars, recesses, etc. In his book Outrage, Vincent Bugliosi faults Ito for allowing the issue of race to enter the trial by permitting F. Lee Bailey's cross-examination of Mark Fuhrman with respect to his use of racial epithets.

During the trial, the prosecution requested that Ito recuse himself when it came to light that his wife, Margaret York, had been detective Mark Fuhrman's superior officer in the past. Fuhrman had been called to testify by the prosecution regarding his discovery of evidence in the case. During cross-examination, Fuhrman claimed that he had not used racial epithets to refer to African-Americans during the last ten years. Simpson's defense team unearthed tapes in which Furhman had used racial epithets as recently as 1988, and they wished to introduce them as evidence to prove that Fuhrman had perjured himself in order to discredit his testimony. However, in the tapes, Fuhrman disparages York's appearance and suggests that she used her gender to advance in the police force. The prosecution requested that Ito step down because they felt that derogatory remarks toward his wife might bias Ito against Fuhrman, though prosecutors later withdrew the request out of fear that it would result in a mistrial.

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