Twenty Drosophila aged 5 10 days were placed on standard medium and allowed to mate for 1 h, after which they were transferred to culture vials containing standard medium plus various concentrations of PA and allowed to lay eggs for 2 h.
Furthermore, they show that sulfamethoxazole (SMX), a sulfa-drug antibiotic and well-known pABA mimic, elicits a dose-dependent lifespan extension when added to C. elegans growth medium containing the standard laboratory E. coli OP50 diet.
Single flies were kept in individual flasks containing standard medium.
For irradiation of first-instar larvae, they were transferred directly after hatching to fresh vials containing standard medium.
Newly eclosed adults were collected, sorted by sex and transferred to fresh vials (10 males or females per vial) containing standard medium and aged at 29°C.
Known numbers of cells were plated into Petri dishes containing standard medium, and colonies >50 cells were counted after 10 days of growth.
For phenotypic observation, ten seedlings were manually transferred to test media containing standard medium alone or also including the indicated additives, and were examined using a dissecting microscope (Leica EZ4 HD).
For experiments in Figure 1, adult flies were allowed to mate for 2 d following eclosion and then collected, sorted by sex, and transferred to fresh vials (≤15 males or females per vial) containing standard medium and aged at 25°.
For the determination of macrocolony morphology, freshly prepared agar plates containing standard LB medium (Miller, 1972) or salt-free LB medium (LBnoS) were inoculated with 5 μl overnight culture applied as small spots.
Seeds were vernalized, planted in pots containing standard planting medium, and germinated in greenhouses near each garden in Spring 2006 prior to being transplanted to the gardens in July 2006.
