ROS production regulates Delta ligand trafficking. (A) ATP measurements from control and PHB1-RNAi egg chambers (n = 3 independent samples). (B) Seahorse-based oxygen consumption rate from control and PHB1-RNAi ovarioles (n = 6 sets of three ovarioles). (C) Measurements of TMRE fluorescence from control and PHB1-RNAi egg chambers (n = 10 independent samples). (D and E) TMRE stained images of control, PHB1-RNAi. (F) ROS measurements from control, PHB1-RNAi, and PHB1-RNAi +SOD2 ovarioles. Follicle cells function as an internal control. The data is expressed as a nurse cell/follicle cell fluorescence ratio. (n = 10 independent samples). (G–I) whole ovary images of control, PHB1-RNAi, and PHB1-RNAi +SOD2 egg chambers (2 copies of GAL4). (J–L) DHE staining images of ROS levels of control, PHB1-RNAi, and PHB1-RNAi +SOD2 egg chambers (1 copy of GAL4). (M–O) Immunofluorescence images of control, PHB1-RNAi, and PHB1-RNAi +SOD2 egg chambers (one copy of GAL4) stained with Delta antibodies. (P and S) DHE staining of control and ND-75-RNAi egg chambers. (Q, R, T, and U) Immunofluorescence images of control and ND-75-RNAi egg chambers stained with Delta antibodies (1 copy of GAL4). 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.