Figure 1. General procedure.
HeLa cells co-expressing a GFP-labeled Golgi localized protein and VSV-G are mixed with HeLa cells expressing a RFP-labeled Golgi localized protein. Cell fusion is triggered by acidic exposure of cell surface targeted VSV-G. Cycloheximide was added 1 hr prior to fusing the cells and during the imaging procedure, to prevent de novo protein synthesis. Golgi-content mixing is assessed by live imaging confocal imaging, visualization and characterization of the putative transport intermediates are assessed by STED microscopy.