Data file: USV preferences raw data- habituation.

Male house mice (Mus musculus) produce complex ultrasonic vocalizations (USVs), especially during courtship and mating. Playback experiments suggest that female attraction towards recordings of male USVs depends on their social experience, paternal exposure, and estrous stage. We conducted a playback experiment with wild-derived female house mice (M. musculus musculus) and compared their attraction to male USVs versus the same recording without USVs (background noise). We tested whether female attraction to USVs is influenced by the following factors: (1) social housing (two versus one female per cage); (2) neonatal paternal exposure (rearing females with versus without father); and (3) estrous stage. We found that females showed a significant attraction to male USVs but only when they were housed socially with another female. Individually housed females showed the opposite response. We found no evidence that pre-weaning exposure to a father influenced females’ preferences, whereas estrous stage influenced females’ attraction to male USVs: females not in estrus showed preferences towards male USVs, whereas estrous females did not. Finally, we found that individually housed females were more likely to be in sexually receptive estrous stages than those housed socially, and that attraction to male USVs was most pronounced amongst non-receptive females that were socially housed. Our findings indicate that the attraction of female mice to male USVs depends upon their social experience and estrous stage, though not paternal exposure. They contribute to the growing number of studies showing that social housing and estrous stage can influence the behavior of house mice and we show how such unreported variables can contribute to the replication crisis.