Fig. 3. Vps35 knockdown affects endocytosis and tagged Vps35 localises to endosomes in S2 cells. (A) Uptake of fluorescent mBSA by cells after RNAi knockdown of novel candidate endocytic proteins. Only knockdowns that significantly inhibited endocytosis and relevant control data are shown; the complete data set is presented in supplementary material Fig. S1. Comparisons are to water control. (B) Single confocal sections showing localisation of Vps35-mRFP (magenta) in S2 cells. Colocalisation with markers of early endosome (anti-Rab5), late endosome (Rab7-EGFP) late endosome and lysosome (spinster-EGFP) and recycling endosome (Rab11 antibody) compartments (green) is shown. Cells were co-transfected with each combination of plasmids for 24 hours and expression was induced with 1 mM CuSO₄ for 4 hours before imaging.