Abl knockdown alters Notch endocytic distribution in S2 cells similarly to abl loss in wings. (A–D″) Co-imaging of Notch and endosomal markers in WT and Abl dsRNA-treated S2 cells. Representative single confocal sections are shown. Transfections: Notch in A and C and Notch + Abl dsRNA in B and D. Stainings: Notch and Eps15 in A and B and Notch and Hrs in C and D. Scale bars = 5 µm. (E) Pearson’s coefficient–based co-localization analysis of Notch and endosomal markers shown in A–D. Each dot represents the Pearson’s coefficient from an individual S2 cell, with an average of 20 evaluated per condition. A t-student means comparison test was performed, and P values <0.05 (*), <0.01 (**), <0.001(***), and <0.0001 (****) are indicated. (F) Relative Notch activity (NRE > luciferase reporter) produced by co-transfection of dsRNAs targeting Notch trafficking regulators, in comparison to co-transfection of LacZ dsRNA or transfection of Notch alone. A t-student means comparison test was performed, and P values <0.05 (*), <0.01 (**), and <0.001(***) are indicated.