Spdo is up-regulated in the absence of dEHBP1. (a–b′′′) Spdo is up-regulated in dEHBP1^−/− ESO cells but not restricted in a Lva-positive Golgi compartment (a–a′′′) and not accompanied by a similar accumulation of Delta (b–b′′′). Arrows indicate to the colocalization of intense Spdo punctae with Delta in both wild-type and mutant cells. (c–c′′′) Spdo is up-regulated in dEHBP1^−/− ESO cells, but not accompanied by accumulation of Notch (d–e′) Notch does not display aberrant localization along the xz axis in ESO clusters, indicated by the asterisks in c′′. (f–f′′′) Endocytosis of Notch is not altered in the absence of dEHBP1. (g–i) Spdo is up-regulated in dEHBP1^−/− ESO cells, but not excluded from the FasIII-positive part of the plasma membrane, as shown in single confocal sections. Magnification of parts of the plasma membrane, included in the boxed regions in g, are shown for a wild-type, GFP-positive ESO cluster (h) and for mutant, GFP-negative, ESO clusters (i). (j) Quantification of the FasII-positive area that is occupied by Spdo in control and dEHBP1^−/− ESO clusters indicates that Spdo is able to reach the plasma membrane in the absence of dEHBP1. Bars, 10 µm.