ROS production regulates Delta protein trafficking. (A) LC/MS GSH/GSSG ratio measurements from control and PHB1-RNAi egg chambers (n = 8 biological samples). (B) LC/MS measurements of Methionine-sulphoxide levels from control and PHB1-RNAi egg chambers (n = 8 biological samples). (C) images of mito-roGFP (green) fluorescence from control and PHB1-RNAi expression intestines. (D) Cytosolic delta levels of control, PHB1-RNAi, and PHB1-RNAi +SOD2 egg chambers (n = 20 independent samples). (E) Cytosolic delta levels of control and ND-75-RNAi egg chambers (n = 20 independent samples). (F) oxygen consumption rate measurements from control, walrus-RNAi, and MTPalpha-RNAi staged egg chambers (N = 6 sets stage 10 egg chambers). Egg chambers were used in this experiment to correct for delays in development. (G) ROS levels from control, walrus-RNAi, MTPalpha-RNAi, and ND75-RNAi egg chambers based on fluorescent imaging. Follicle cells are used as an internal control, and the data is expressed as a ratio of nurse cell/follicle cell fluorescence (n = 20 egg chambers). (H) ROS-level quantification from control and ND75-RNAi egg chambers (n = 20 egg chambers). (I and J) Delta staining images of ovarioles from Walrus-RNAI and MTPalpha-RNAi females. (K) Delta staining of PHB-RNAi+catalase, PHB-RNAi+SOD1, and DAAO-expressing ovarioles. Arrows point to areas where Delta localization to the membrane is rescued. (L) Delta staining images from ND75-RNAi and ND75-RNAi+SOD2 ovarioles. (M) A summary graph measuring the %rescue of ovarioles showing normal Delta membrane staining of all UAS-transgenes used in this study. The expression of all the transgenes used in this graph is driven by matα-GAL4. Each data point represents an experiment of at least 25 ovarioles. Student’s t test was used for all pairwise comparisons, and one-way ANOVA was used for all experiments containing >2 sample groups. Error bars represent the standard deviation.