All tissues were paraffin embedded and derived from males and were cut at 5 μM.
For testing courtship and copulation of a male and female mixed population, the wings of either males or females were cut off on ice to distinguish the sex 1 h before ethanol exposure.
The three individual samples of 109 bp length (one territorial male, one sneaker male and one female) were cut down to 48 bp to avoid a length bias in the expression value calculation, as the other 12 individual samples (four territorial males, four sneaker males and four females) were sequenced at a shorter length.
Male and female antennae were cut at the base and fixed in glutaraldehyde.
Briefly, three 2-day-old males, isolated at L4 stage, were cut at the posterior portion with a needle in 20 μl sperm media (50 mM Hepes, pH7.0, 45 mM NaCl, 25 mM KCl, 1 mM MgSO4, and 5 mM CaCl2) freshly supplemented with polyvinylprolidone (PVP) 40,000 molecular weight (Sigma) and the activator pronase (Roche, Indianapolis, IN) at the final concentration of 10 mg/ml and 500 μg/ml on a slide.
Frozen brains of young (10 12 weeks of age; 300 350 g body weight) male Sprague-Dawley rats (Harlan, Netherlands) were cut at −12 °C into sagittal slices 20 μm thick using a Leica microtome, and the slices were thaw-mounted on Superfrost (70 × 22 mm, Fischer) adhesive slides.
Sugar also wrote several lines for Marshall Lee Marcelinene's male counter-part — but they were cut for time.
The gonadal tissues (testis for male and ovary for female) were cut into cubes of approximately 0.5 × 0.5 × 0.5 cm in size and immediately immersed in liquid nitrogen overnight, and then stored at −80 °C until RNA extraction.
Frozen males were decapitated and their heads were cut at both sides to open up the postpharyngeal gland, which is the storage organ of the male sex pheromone [ 40, 57].
Next came an uncomfortable couple of days where I had to ask my male friends whether or not they were cut, which apparently isn't the most normal thing to text someone at 9 AM on a Tuesday.
