Fluorescence patterns of GFP/Cherry-tagged E-Cad and Spdo. (A–A″) Fluorescence signals produced by CadGFP (green) and CadCherry (red) in living pupae showing that GFP mostly revealed the junctional pool of E-Cad, whereas the strongest Cherry signal was detected at endosomes. Note that the junctional pool of E-Cad was also weakly Cherry positive. (B) Domain structure of Spdo showing the four transmembrane segments (black) and the LHELL and NPAF trafficking signals (purple; Tong et al., 2010; Upadhyay et al., 2013). SpdoGFP1 was described previously (Couturier et al., 2013). The points of GFP (green), Cherry (red), and GFP-Cherry tandem dimer (green and red) insertion are indicated (in amino acids). The results of the genomic rescue experiments are shown on the right. (C) spdo mutant flies rescued by one copy of the SpdoCherry2GFP3 BAC. (D–D″) Live imaging of SpdoCherryGFP (GFP [green] and Cherry [red]) showing that GFP and Cherry revealed distinct pools. Bars: (A) 10 µm; (D) 2 µm.