Analyzing Drosophila Wing Size and Fertility

Lethal phase analysis was performed as described previously (Vasudevan et al., 2020 (link)). Right wings were severed from 1- to 4-day-old flies and imaged using a Nikon SMZ1500 microscope outfitted with a Nikon 8MP camera with NIS-Elements software. Wing size was measured using the regions of interest feature in ImageJ software.
Female fertility was quantified by placing five 1- to 4-day-old virgin females with five yw males (or Dj-GFP males for fertilization assays) in a vial containing standard medium enhanced with yeast to encourage egg laying. After allowing 1 day for acclimatization, the flies were moved to a new vial and the number of eggs laid in a 24-h period were counted and quantified. Eggs were imaged for Fig. 6B by placing them on an apple juice plate and capturing them with a Nikon SMZ1500 microscope outfitted with 8MP Nikon camera controlled by NIS elements software. Ovaries from female flies in this experiment were dissected in ice-cold PBS and similarly imaged on apple juice plates for Fig. S4A.