Mutations in the ANK domain impair Notch traffic. Wing pouch images from third instar discs expressing CeN (A–A2), CeN-ΔRANK (B–B2), CeN-DM1. (C–C2) and CeN-DM2 (D–D2) under the control of dpp-Gal4. All are derivatives of CeN; CeN-ΔRANK is a deletion of the RAM and ANK domains of the intracellular domain, whereas CeN-DM1 and CeN-DM2 are point mutations in the fourth and fifth ANK repeats (for details see Materials and Methods). All images were taken from the apical level of the wing pouch; the arrows point to the dorsal-ventral (DV) boundary; the eGFP fluorescence (from the CeN molecule) is shown in all cases. (A2–D2) are confocal optical z-sections through the wing pouches. Note that there is an apical accumulation of the CeN mutant molecules in apical levels, particularly clear in the CeN-ΔRANK and fewer vesicles in basal levels, which tend to be of larger size. These are images from a steady state, for pulse chase of one example see Figure S12. (7.86 MB TIF)