Df(3L)Ilp2-3, Ilp5¹ mutants show typical circadian rhythmicity under 12:12 hour light:dark (LD) and constant dark (DD) conditions. However, in the mutants day activity is significantly increased, whereas night activity is significantly reduced, a pattern that is maintained as the flies age. Wakefulness (average activity per awake minute) is not significantly altered, suggesting that Df(3L)Ilp2-3, Ilp5¹ mutants have a greater number of active periods during the day.

At all ages tested, Df(3L)Ilp2-3, Ilp5¹ mutants sleep more at night and less by day than controls. In addition, they have fewer waking periods, and hence sleep bouts, during both day and night, and longer night sleep bouts than are seen in controls.

The increase in sleep fragmentation with age seen in control flies is not observed in Df(3L)Ilp2-3, Ilp5¹ mutant flies. Night sleep duration does not change, day sleep duration also does not change, and day and night sleep bouts do not increase with age.

Unlike in controls, treating Df(3L)Ilp2-3, Ilp5¹ mutant flies with tolbutamide does not increase daytime activity.

Octopamine levels are increased in head extracts from Df(3L)Ilp2-3, Ilp5¹ mutant flies. Levels of tyramine, the precursor of octopamine, are significantly reduced.

Feeding flies with mianserin hydrochloride, an inhibitor of octopaminergic signalling, suppresses the increased day activity, sleep and sleep bout number, but not the night sleep phenotypes seen in Df(3L)Ilp2-3, Ilp5¹ mutants. The increase in lifespan is unaffected.

Unlike in controls, feeding Df(3L)Ilp2-3, Ilp5¹ mutant flies with the TOR inhibitor rapamycin has no effect on the night activity of sleep phenotypes. Feeding older flies (42 days) with rapamycin suppresses the sleep fragmentation phenotype. It results in increased night sleep duration, fewer night sleep bouts and increased bout length, without affecting daytime activity behaviors.

The levels of dopamine biosynthetic enzymes and total dopamine are normal in Df(3L)Ilp2-3, Ilp5¹ mutant flies. However after treatment with rapamycin levels of dopamine transporter (DAT) increased. The reduction in nighttime activity and increase in sleep seen in wild type flies in response to the dopaminergic signalling inhibitor 3-Iodo-L-tyrosine (3IY) is not seen in Df(3L)Ilp2-3, Ilp5¹ mutants, which instead show a slight increase in activity compared to controls. 3IY treatment has no effect on daytime activity levels in either wild type or Df(3L)Ilp2-3, Ilp5¹ mutant flies. The increase in both daytime activity sleep seen in wild type flies in response to methamphetamine is not seen in Df(3L)Ilp2-3, Ilp5¹ mutant flies, whereas nighttime activity increases to levels similar to those of treated controls.

AkhR¹ suppresses the increase in daytime activity seen in Df(3L)Ilp2-3, Ilp5¹ mutants. Night activity, night sleep duration and number of night sleep bouts are similar to controls. The increase in octopamine levels seen in Df(3L)Ilp2-3, Ilp5¹ mutant head extracts is also rescued by AkhR¹.

Dop1R1^f02676 does not suppress the activity and sleep defects seen in Df(3L)Ilp2-3, Ilp5¹ mutant flies.

Scer\GAL4^Cg.PA-mediated expression of Myc^{Scer\UAS.T:Ivir\HA1} does not result in large wings in a Df(3L)Ilp2-3, Ilp5¹ background.

Df(3L)Ilp2-3, Ilp5¹ mutants are 100% viable (females)/60% viable (males); show a severe delay in egg-to-adult development time; females show a severe reduction in adult body weight; have normal median lifespan; show a significant reduction in lifetime fecundity; show increased paraquat resistance; show normal starvation resistance; and have significantly increased lipid and glycogen storage. These phenotypes are not further negatively affected when Ilp7¹ is introduced into this background.

While Df(3L)Ilp2-3, Ilp5¹ homozygotes do not show an increased life span, Df(3L)Ilp2-3, Ilp5¹ heterozygotes are slightly long-lived, while Wolbachia-infected Df(3L)Ilp2-3, Ilp5¹ mutants are extremely long lived.

Df(3L)Ilp1-4, Ilp5¹ mutants are less than 100% viable; show a severe delay in egg-to-adult development time; and females show a severe reduction in adult body weight. These phenotypes are not further negatively affected when Ilp7¹ is introduced into this background.

The dietary restriction response of Df(3L)Ilp2-3, Ilp5¹ females is 'right-shifted' - mutant flies are shorter-lived compared to controls on low yeast concentrations, but longer-lived on high yeast concentrations. Also, whereas wild type flies show a strong increase in egg production between 1x and 2x food (62-81%), Df(3L)Ilp2-3, Ilp5¹ mutants only lay 26% more eggs on the higher yeast concentration.