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Fig. 3. Akhr mutants are starvation resistant. All experiments were performed on 1-week-old male flies, except when specifically noted otherwise. Throughout the figure, data for Akhr^null flies and Akhr^rev flies are represented in blue and yellow, respectively. (A) Starvation resistance of Akhr mutant flies. Starvation resistance profiles were quantified on the Trikinetics Drosophila Activity Monitoring System (DAMS), with individual monitoring tubes containing 2% agarose, but no other food source. The time of death of an individual male fly was defined to be the time of last recorded locomotor activity, which correlated well with starvation profiles obtained from direct observation. Average starvation resistances are given for each genotype (N=16) with their corresponding standard deviations. Akhr^rev flies have starvation resistance that is comparable to y¹w^67c23 flies, the genetic background from which the Akhr^p line was generated. Akhr^null flies were markedly starvation resistant when compared to Akhr^rev control flies, and Akhr^p flies showed an intermediate phenotype (Student's t-test, ^*P<0.05 for both comparisons). Starvation resistance comparisons between genotypes were repeated at least three times. (B) Both young and older (1 week) Akhr^null flies have enhanced starvation resistance when compared to age-matched Akhr^rev control flies (Student's t-test, ^*P<0.05). (C) Akhr^null flies are able to mobilize triglyceride stores, as reflected by the decreased lipid levels of flies starved for 72 h. (D) Akhr mutants do not have defective locomotor activity or circadian rhythm. Average number of midline crossings (N=16 for each genotype) were recorded every 30 min for fed Akhr^rev and Akhr^null using the DAMS. No gross defects in locomotor activity or circadian rhythm were observed in Akhr^null flies. (E) Total locomotor activity (counted as number of midline crossings) for the first 24 h of starvation for each genotype, with corresponding standard deviations (N=16 for each genotype). The differences in locomotor activity between all starved lines were not statistically significant.

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