Fig. 5

Rab4b-dependent retrograde trafficking involves the GARP complex. A Schematic representation of VPS52 structures. The blue circles represent the coiled-coil domain. B Schematic representation of the interaction detected in the yeast two-hybrid system, performed as in [26, 27], between full-length VPS52 and the indicated forms of Rab4b or lamin as negative control. +  +  +  +  + is for an interaction is detectable before 30 min.—no detectable interaction after 24 h. (C) Quantification of iCD8-M6PR in the Golgi area of cells overexpressing HA-Rab4b Q67L (empty circles) and in the surrounding non-transfected cells (NT, filled circles) according to protocol 1 (Additional file 1: Fig. S2A). Each dot represents one cell. Each color (black, blue, or green) indicates independent experiments. ***,**** indicates that the differences are significant compared to NT siCtrl with and ^#### indicates that the difference is significant compared to NT siVPS54 with p < 0.001 or p < 0.0001 (one-way ANOVA). The significance is the same whether the 3 experiments are analyzed together or independently. D Representative images corresponding to the quantification shown in (C), stars are for transfected cells. The downregulation of VPS52 is validated in an independent well in each experiment by Western blot or immunolabeling as shown in (Additional file 1: Fig. S3A-B)