Relative florescence intensity (RFI) of proliferating cells in in vitro cultured midguts dissected from insects injected with Tcbr or malE dsRNA exposed to DMSO, 20E, JH III or 20E + JH III and quantified using BrdU labeling. RFI was measured using Olympus Flouview software version 1.5. Squares of constant area (6662mm2) and length (26 mm) were drawn on composite Z-stack images. The average intensity in the marked area against the background was measured using the software. All other parameters (PMT, Gain, Offset, zoom) were the same for each image documented. Mean ± SE of three independent experiments (n = 15) are shown.