The present study is one of few to investigate both anti-trans discrimination and anti-trans prejudice. It examined four individual factors (religiosity; political beliefs; affiliation with LGBTQ [lesbian, gay, bisexual, transgender, and queer] people; gender role beliefs) through a lens of hetero-cis-normativity to understand their association with anti-trans attitudes and reported behaviors. Using a sample of 302 cisgender college students from across the United States, hierarchical multiple regressions on bootstrap samples were used to analyze how these factors are associated with anti-trans attitudes and behaviors. More liberal political beliefs, affiliation with more LGBTQ friends and family members, and less traditional gender role beliefs were related to more positive attitudes toward transgender people. Less traditional gender role beliefs and more positive attitudes were associated with more positive reported behaviors toward transgender individuals. Interventions designed to challenge traditional gender role beliefs and approximate affiliation with LGBTQ persons may be most effective to reduce pervasive hetero-cis-normative prejudice and discrimination within schools.