Figure 7.

Endocytic pathway as a source of HSV envelope and experimental evaluation. (A) 1. Synthesis of viral glycoproteins in the endoplasmic reticulum. 2. Post-translational modifications of viral glycoproteins through Golgi apparatus. 3 Glycoproteins arrived at trans Golgi network and transport to the plasma membrane. 4. Glycoproteins arrive at the plasma membrane exposed to the extracellular space. 5. Staining of viral glycoproteins with primary antibodies and endocytosis. 6. Transport of antibody-stained viral glycoproteins through early endosomes. 7. Antibody-stained glycoprotein-containing tubules wrap cytoplasmic capsids forming virions with a double membrane. Cell fixing and secondary antibody fluorophore-conjugated staining for confocal microscopy evaluation. ER, endoplasmic reticulum; GA, Golgi apparatus; PM, plasma membrane; TGN, trans Golgi network; EE, early endosomes. (B) In order to study the process of virion maturation in the cytoplasm, it is possible to perform an antibody uptake assay. First, a confluent plate of cells is infected, 1 h after the cells are washed with acid buffer in order to inactivate residual viral particles. Then the cells are washed with PBS and incubated in free bovine serum medium with anti-glycoprotein primary antibodies for 16 h. Then, the cells are fixed and stained with a secondary antibody conjugated with a dye for immunofluorescence detection.