Extended Data Fig. 7: Activation of song pathway neurons.

a, Solitary ATR-fed P1a males produce song when exposed to the same LED stimulus used in Figure 4. In solitary males, song production is both long-lasting and time-locked to the LED stimulus. Number of animals in parentheses (n=5), heavy line represents mean, shading represents SEM. b, ATR-fed P1a males courting a female produce significantly more Pfast (p = 1e-4), Pslow (p = 1e-3), and significantly-different amounts of sine song (p = 0.009). All p-values from Mann-Whitney U-test. Animals from (a) (n=5), center lines of box plots represent median, the bottom and top edges represent the 25th and 75th percentiles (respectively). Whiskers extend to +/- 2.7 times the standard deviation. c, The probability of observing each song mode aligned to the opto stimulus shows that LED activation of flies not fed ATR does not increase song production. Number of animals in parentheses, heavy line represents mean, shading represents SEM. d, The probability of the model being in each state aligned to the opto stimulus shows that LED activation of flies not fed ATR does not change state residence. Error bars represent SEM. Number of animals in parentheses, heavy line represents mean, shading represents SEM. e-f, ‘Opto’ filters represent the contribution of the LED to the production of each type of song for (e) ATR+ and (f) ATR- flies. Number of animals in parentheses, heavy line represents mean, shading represents SEM. The filters for each strain and song type are not significantly different between states. g, Measuring the maximal change in state probability between LED ON and LED OFF shows that only pIP10 activation produces a significant difference between ATR+ and ATR- flies (two-tailed t test). Number of animals in parentheses in (e-f), center lines of box plots represent median, the bottom and top edges represent the 25th and 75th percentiles (respectively). Whiskers extend to +/- 2.7 times the standard deviation. All p-values from two-tailed t test.