Fig 2. Quantitative analysis of wing disc shapes and nuclear positions along the major axis of imaginal wing discs at 96 hours AEL.

(A) Representative pre-processed image of a wing disc cross-section stained with DAPI (nuclei) and phalloidin (staining F-actin). These images were used segment nuclei and surfaces defining the wing disc shape. B) The fractional position between the apical and basal surface serves as a metric for quantifying relative nuclear positions. Note: heights of columnar pouch cells were calculated as the sum of L_A and L_B. C) Representative segmentation of nuclei. Blue and red lines correspond to L_A and L_B in B, respectively. D) Basal surface segmentation for multiple samples in different colors (N = 13). E) Composite representation of wing disc morphology generated from samples in D. F) Quantification of global and local curvatures. Fig A and Fig B in S4 Text provide additional details on the image processing pipeline. G) Representative image highlighting average curvature quantification used for H, the curvature was found using a circumcircle approach. H) Curvature quantification of regions of interest along the pouch as well as intensity quantification of integrin along the basal side of the pouch.