Fig. 4

Activating noradrenergic transmission with isoproterenol and phenylephrine marginally depresses caffeine-induced c-fos expression. a Sleep traces for larvae exposed to combinations of 10 µM isoproterenol + 10 µM phenylephrine (“Iso + Phenyl”) and/or caffeine. At top left, the pale blue horizontal bar shows the isoproterenol + phenylephrine exposure window while the gold bar indicates the presence of caffeine. The post-drug rebound sleep period of a is summarised for each larva in b (upper chart). Lower chart shows the effect size (with 95% CI) of Iso + Phenyl treatment among water-treated and caffeine-treated groups. c qRT-PCR on groups of ~ 18 larvae reveals that each group of larvae pre-treated with isoproterenol + phenylephrine and then caffeine (n = 3 biological replicates) had lower relative c-fos expression than the groups of larvae treated with water and then caffeine (n = 3 biological replicates); see also Fig. S5a. Each square is the mean of three technical replicates. d The average relative c-fos expression induced by each condition is strongly positively correlated (R2 = 0.889) with the total rebound sleep that was induced by the same drug condition (from a and b)