Lifespan Analysis of Mated Flies

Lifespan measurements were conducted in three cohorts (replicates) of ∼80 mated males and 80 mated female flies of each genotype (n = 240) separately. These flies of a specific sex and genotype were kept separately in round bottom polypropylene bottles (8 oz, Genesee Scientific, San Diego, CA, United States) inverted over 60 mm Falcon Primaria tissue culture dishes serving as lids (Becton Dickinson Labware, Franklin Lakes, NJ, United States) containing diet (15 mL). The diet dishes were replaced on alternate days after tapping flies down to the bottom of the bottle. Mortality was scored daily, and Kaplan–Meier survival curves were generated and Log-rank (Mantel–Cox) tests were conducted using GraphPad Prism (v 5.01). The Gompertz–Makeham maximum likelihood estimates (MLE) were conducted using WinModest (v.1.0.2) (Krishnan et al., 2009 (link)).