AbstractIncubation of cultured cells in hypertonic medium and sodium‐free medium have been shown to block transport at two different stages along the endocytic pathway. To determine the effects of these treatments on the exocytic pathway, we studied the transport of the membrane glycoprotein of vesicular stomatitis virus (VSV‐G) in cells infected with tsO45 mutant virus. This mutant synthesizes a VSV‐G that accumulates in the endoplasmic reticulum (ER) when cells are incubated at 39.5°C. In addition, VSV‐G accumulates in the post‐ER pre‐Golgi compartment when cells are incubated at 15°C and in the trans‐Golgi network (TGN) when cells are incubated at 18°C. Upon transfer of cells to 32°C in control medium, VSV‐G exits each of these compartments and is transported to the cell surface. Incubation in sodium‐free medium at 32°C did not block transport from any of these three compartments. In contrast, incubation in hypertonic medium blocked export from the ER, transport from the pre‐Golgi compartment to the Golgi complex, and transport from the TGN to the cell surface. Our results, in combination with previous studies, suggest that hypertonic medium blocks at least five distinct transport steps: the three exocytic steps described here, endocytosis from the cell surface, and transport of cell surface proteins into the Golgi complex. This raises the possibility that vesicular transport in different parts of the cell shares common elements that are inh