Loss of abl increases Notch levels and endocytic accumulation. (A–B″) Maximal projections showing Notch protein expression detected by indirect immunofluorescence with anti-Notch in 32 h APF WT and abl^null pupal wings, with zoomed views of the PCV region in A′, A″, B′, and B″. Scale bars = 100 µm (A and B), 20 µm (A′ and B′), or 5 µm (A″ and B″). Additional, and partially overlapping with (A″ and B″), zoomed views of the wings in A and B are shown in Fig. S2, A and B. (C and D) Plots of Notch fluorescent intensity values across the anterior-posterior axis of the wing blade (yellow lines in A and B). (E) Quantification of Notch fluorescent intensity at the cell cortex and cytoplasm of vein and intervein cells in 32 h APF wings. Each dot represents the individual measurement of a cell border (N = 158 in WT vein areas, N = 156 in abl vein areas, N = 130 in WT intervein areas, and N = 129 in abl intervein areas) or a cell cytoplasm (N = 70 in WT vein areas, N = 80 in abl vein areas, N = 52 in WT intervein areas, and N = 53 in abl intervein areas). A t-student means comparison test was performed, and P values <0.05 (*), <0.01 (**), <0.001(***), and <0.0001 (****) are indicated. (F and G) Quantification of the area and fluorescent intensity of Notch⁺ subcellular foci, respectively. Each dot represents the individual measurement of a cytoplasmic Notch⁺ structure (N = 455 in WT vein areas, N = 535 in abl vein areas, N = 296 in WT intervein areas, and N = 355 in abl intervein areas). A t-student means comparison test was performed, and P values <0.05 (*), <0.01 (**), <0.001(***), and <0.0001 (****) are indicated. (H) Quantification of the frequency of Notch⁺ subcellular foci. Each dot represents the average number of Notch⁺ structures per cell from measurements of 28 × 28-µm regions, as shown in 2A″ and B″, S2 A and S2 B. N = 20 regions were measured for each condition. The average cell number per region was 66.75 (WT vein), 64.8 (abl vein), 53.55 (WT intervein), and 56.2 (abl intervein), respectively. A t-student means comparison test was performed, and P values <0.05 (*), <0.01 (**), <0.001(***), and <0.0001 (****) are indicated.