Quantification of UAS‐eYFP signals along the profile of an intestine

Intestines were stitched together using the stitching tool in ImageJ to show entire intestine profile in the eYFP channel. The image was then made into a maximum projection, flattening all z slices into one image. The line tool was then used at a width of 500 pixels (approximately the width of the average intestine), and a line was drawn along the entire profile of the intestine. From there, the plot profile analysis was conducted in ImageJ, providing the average intensity value of eYFP signals across the 500 pixel width at each pixel along the intestinal profile (approximately 10,000–20,000 measurements per intestine). These measurements were then used to provide summary values for each intestine. Every 1,000‐pixel measurements were averaged to provide one value, giving 10–20 values per intestine. These averaged values were then combined from at least 10 individual intestines into a single dataset.