Prejudice towards trans women is an under-examined but a critical social problem in Turkey. Patriarchal femininity sanctions women who do not comply with the idealised female bodies and feminine norms. On the other hand, ideals of hegemonic masculinity necessitate that men avoid femininity. In that sense, prejudice against trans women ostensibly stands at the intersection of hegemonic masculinity ideology and femininities that diverge from patriarchal feminine norms. Therefore, this study examines the mediatory effect of femmephobia in the link between masculinity ideology and prejudice towards trans women as measured by negative affect and social distancing motivations. Results indicate that femmephobia significantly mediates the relationship between the endorsement of masculinity ideology and prejudice towards trans women. Among three mandates of masculinity ideology, the endorsement of the antifemininity mandate was the most solid indirect predictor of negative affect and social distance through femmephobia, followed by mandates of toughness and status, respectively. Although participant gender does not have an indirect effect on the outcome variables, it has a direct effect on men's social distancing intentions but not on negative emotions. The results are discussed in the light of existing literature which intersects with the devaluation of femininity.