Fig. 12

The HSV-1 gB co-localizes with Gag-GFP at the cell surface. COS-7 cells were transfected with either the vector expressing gB or co-transfected with gB and Gag-EGFP. At 24 h, cells on cover slips were washed, fixed, permeabilized and blocked as described above. The cover slips were reacted with a mouse monoclonal antibody against HSV-1 gB and an appropriate secondary antibody and counterstained with DAPI (1 μg/ml) for 5 min as described in the Materials and Methods section. The cover slips were mounted and examined using a Leica TCS SPE confocal microscope using a 63 × objective with a 2 × digital zoom using the Leica Application Suite X (LAS X, LASX) software package. A 405 nm filter used to visualize the DAPI, 488 nm filter was used to visualize the Gag-GFP, and a 594 nm filter to visualize gB staining. a–d Cells transfected with vector expressing gB. e–h Cells co-transfected with vectors expressing gB and Gag-GFP. a, e Visualization of Gag-EGFP. b, f Visualization of gB staining. c, g Visualization of DAPI staining. d, h Merge of the three panels to the left. The inset below Fig. 11h is a region of the cell plasma membrane showing co-localization of Gag-EGFP and gB. The results shown are representative of examining 50 transfected cells