Nedd4Lo promotes Notch internalization into the LE lumen to attenuate signaling. (A–B″) Single confocal sections of representative S2 cells showing Notch (indirect immunofluorescence) and Rab7-EYFP. “n” marks the cell nucleus. Transfections shown: (A) Notch + Nedd4S and (B) Notch + Nedd4Lo. Scale bars = 5 µm. (C) Plot showing profiles of Notch levels across LE diameters in A and B. Sample size (# of LEs evaluated): Notch (WT, N = 38), Notch + Nedd4S (N = 27), and Notch + Nedd4Lo (N = 36). (D) Relative Notch activity in manipulations in A and B. A t-student means comparison test was performed, and P values <0.05 (*), <0.01 (**), and <0.001(***) are indicated. (E and F) en > Nedd4S and en > Nedd4Lo adult wings, respectively. Scale bars = 100 µm.