The existence of ovulatory-cycle shifts in women’s mate preferences has been a point of controversy. There is evidence that naturally cycling women in their fertile phase, compared with their luteal phase, evaluate specific behavioral cues in men as more attractive for sexual relationships. However, recent research has cast doubt on these findings. We addressed this debate in a large, preregistered, within-participants study using salivary-hormone measures and luteinizing-hormone tests. One hundred fifty-seven female participants rated the sexual and long-term attractiveness of 70 men in dyadic intersexual interactions in natural videos. Multilevel comparisons across two ovulatory cycles indicated that women’s mate preferences for men’s behaviors did not shift across the cycle for either competitive or courtship behavior. Within-women hormone levels and relationship status did not affect these results. Hormonal mechanisms and implications for estrus theories are discussed.

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