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Homophobia and biphobia negatively impact the mental health of gay, bisexual, trans, Two-Spirit, and queer men and non-binary individuals (GBT2Q) and sexual and gender minority men, but little is known about the impact of gender-related oppression. The current study examines the impact of pressure to conform to masculine norms in Canada-based GBT2Q individuals. Specifically, the associations between (a) gender expression and pressure to be masculine and (b) pressure to be masculine and depression, anxiety, and self-rated mental health were investigated. Drawing from an online national cross-sectional survey of 8,977 GBT2Q individuals and sexual and gender minority men living in Canada aged 15 years or older, 56.4% ( n = 5,067) of respondents reported experiencing pressure to conform to masculine norms. Respondents were more likely to report masculine pressure if they were younger than 30 years, described their gender expression as fluid, identified their sexuality as queer, were an ethnoracial minority, and were trans. Pressure to be masculine was associated with increased odds of depression, anxiety, and reporting poor or fair mental health. The current study provides evidence of the detrimental impact of pressure to conform to masculine norms on the mental health of gay, bisexual, trans, Two-Spirit, and queer men and non-binary peoples., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.