Ovulatory cycle shifts in women's mate preferences have been documented for several physical and behavioral traits. Research suggests that, at peak fertility, women tend to prefer men with characteristics that reflect good genes for short-term sexual relationships. However, existing findings have been criticized for methodological flexibility and failing attempts to replicate core results. In a large (N = 157), pre-registered, within-subject study spanning two ovulatory cycles, we investigated cycle shifts in women's mate preferences for masculine bodies. Using a large set of natural stimuli, we found that when fertile, women's ratings of male bodies increased for sexual as well as for long-term attractiveness. Both effects were partially mediated by the estradiol-to-progesterone-ratio. Furthermore, moderation analyses revealed that both shifts were only evident in women in relationships, but not in singles. Contrary to previous findings, male masculine traits did not interact with cycle phase to predict attraction, indicating that women's preferential priorities do not shift. Taken together, our results do not support women's mate preference shifts, as assumed by the good genes ovulatory shift hypothesis, but are consistent with shifting motivational priorities throughout the cycle. Implications of these results for female estrus theories and methodological recommendations for future research are discussed.