Fig. 4: Female copulation song decreases remating.

a Experimental design for testing the effect of female song. Data from the playback condition is shown in red, from the control silence condition in blue. b Copulation duration of mute females with wild type males, with and without female song playback. c Adult progeny after 6 days of egg laying from copulations of mute females with wild type males, with and without female song playback. d Number of sperm in female storage organs after copulations with and without female song playback. e Experimental design for testing the effect of female song playback under subsequent reproductive competition. The first male and it’s progeny is displayed in orange, the subsequent males and their progeny in magenta. f Female remating after copulations with and without song playback. *p = 0.0258 (day 1), **p = 0.0097 (day 2), *p = 0.0102 (day 3), *p = 0.0446 (day 4). Fisher’s exact test (two-sided). g Adult progeny after 6 days of egg laying, from copulations of mute females with wild type males, with and without female song playback, with the female subsequently housed with 3 males carrying genetic markers Sb/ Tm3, Ser. h Relative reproductive advantage of the first male ((progeny first male−progeny subsequent males)/all progeny)) in the experiment outlined in e after 6 days of egg laying. n is indicated after the conditions, error bars indicate standard error of the mean, **p = 0.0099, permutation testing (one-sided). i Percentage of wild type females muted by wing amputation remating within 20 h with wild type males after copulations with and without song playback. *p = 0.014, Fisher’s exact test (two-sided). j Latency to the first remating after copulations with and without song playback. **p = 0.0035, Mann-Whitney test (two-sided). k Schematics of proposed copulatory dialog. In b, c, d, g and j each data point represents one couple, n is indicated after the conditions, error bars indicate median and interquartile range, no significant difference in b, c, d and g, Mann-Whitney test (two-sided). Source data are provided as a Source data file.