O Brother, Where Art Thou? does not feature any identifiable LGBTQ+ characters or themes. The narrative is entirely focused on the adventures of three escaped convicts in the American South during the Great Depression, with no elements touching upon queer identity or experiences.

O Brother, Where Art Thou? does not include any identifiable transsexual characters or themes. The narrative focuses on the adventures of three escaped convicts in the Depression-era South, with no elements pertaining to transgender identity or issues.

The film does not feature any female characters engaging in or winning direct physical combat against male opponents. Female characters are present but do not participate in action sequences that meet the specified criteria.

The film is a loose adaptation of Homer's Odyssey, which does not define characters by modern racial categories. All major characters are portrayed consistently with their setting and the general understanding of their source material counterparts, or are new characters whose race aligns with their inspiration. No character established as one race is portrayed as another.