ROS regulates endosomal recycling by reducing the levels of PIP2 in membranes. (A) Images of ovarioles from control and PHB-RNAi females. Top image DAPI, middle Images PIP2, bottom image LUT intensity image of PIP2 levels. (B) images of (MCF7) and (BT549) cells stained with DAPI and PIP2 antibodies (LUT intensity image of PIP2 levels). (C) Images of control (3T3) and (BT549) cells stained with PIP2 antibodies (LUT intensity image of PIP2 levels). Fluorescence levels are expressed with a Fire LUT. Arrows indicate a PIP signal in the cell body. (D) Quantification of PIP2 antibody staining levels from control and PHB-RNAi ovarioles. (E) Quantification of PIP2 antibody staining fluorescence from (MCF7) and (BT549) cells. (F) Quantification of PIP2 antibody staining fluorescence from control (3T3) and (BT549) cells. (G) Quantification of the percentage of follicles containing punctate Delta staining from three independent experiments. (Each experiment contained at least 25 egg chambers, and the total number of eggs assayed for each group was at least 85). (H and I) Delta antibody staining images from control and PIP5K-RNAi (sktl-RNAi) ovarioles. Arrows point to Delta puncta. (J and K) peb antibody staining images from control and PIP5K-RNAi (sktl-RNAi) ovarioles. (L) Quantification of the percentage of follicles containing abnormally low peb expression from three independent experiments. (Each experiment contained at least 25 egg chambers, and the total number of egg chambers assayed for each group was at least 85). 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.