Fig. 6: Ihog at membranes is withdrawn by endocytosis and degradation.

A–D Crops from a lateral section of hh.Gal4, tub.Gal80^ts > UAS.RHGmiRNA wing discs expressing UAS.Ihog^RFP (A), UAS.Ihog^CFP and UAS.EGFR^RNAi (B), UAS.Ihog^RFP and UAS.Shi^RNAi (C) or UAS.Ihog^RFP plus EGFR^RNAi, together with UAS.Shi^RNAi to block endocytosis (D). Note a severe increase in puncta numbers after EGFR downregulation. Note also that Ihog^RFP levels at plasma membranes decrease after inhibition of EGFR but are significantly rescued when endocytosis is simultaneously inhibited. Scale bars = 10 μm. E–F1) Incubation with Dynasore, inhibitor of the GTPase activity of Dynamin, at 25 °C for 40 min, prior to incubation with 3.7 mM Dextran−647 for 10 min. Note the increases Ihog^CFP in the plasma membrane after UAS.EGFR^RNAi expression (E, F). Note also the absence of Dextran labelled vesicles (E1, F1). G Vesicle-like puncta were quantified within a defined area for the three treatment conditions. A total of 8 wing discs per treatment were analysed, and a ANOVA test was performed, yielding p-value < 0.0001. H, I) Crops of lateral sections of hh.Gal4, tub.Gal80^ts > UAS.RHGmiRNA wing discs expressing Ihog^CFP as control (H) and UAS.Ihog^CFP together with UAS.EGFR^RNAi (I) and incubated with 100 nM Lysotracker (green) for 40 min at 25 °C and for the last 10 min with 3.7 mM Dextran−647 (red). Scale bars = 10 μm. The split channels in the amplified insets show in more detail the increase of Dextran and Lysotracker vesicles after EGFR^RNAi expression. Scale bar = 5 μm. Source data are provided as a Source Data file.