Fig 5. Effect of blockade of gK interaction with SPP using gK4 on gK localization in HSV-1 infected cells.

A) Detection of HSV-1 gK in infected cells. Vero cells grown to confluency on chamber slides were infected with 1 PFU/cell of VC1 virus in the presence of 20 μg/ml of dTatgK4 or dTat peptide for 1 hr. After 1hr infection, the media was replaced with fresh media containing each peptide for 16 hr. Infected cells were fixed and immunostained using anti-V5 (for gK) with anti-GM130 (for Golgi), and chicken anti-goat AlexaFluor 488 (for gK) with chicken anti-rabbit AlexaFluor 594 (for GM130) as secondary antibodies. DAPI was used for nuclear staining (blue). There were fewer gK positive cells in the dTat-gK4 treated group than in control dTat peptide-treated group. gK protein colocalized with Golgi protein GM130 in the control peptide group, indicating enrichment of gK protein in the Golgi apparatus. However, gK protein did not colocalize with GM130 in the dTat-gK4 treatment group. gK protein localization to the Golgi was inhibited by blocking the binding of gK to SPP. Photomicrographs are shown at 630X direct magnification. Experiments were repeated twice; and B) Quantification of photomicrographs from A. Different areas of 3 slides/peptide from IHC described above were imaged and the number of HSV-1 gK⁺ cells was counted. Each point represents the mean ± SEM of HSV-1 gK⁺ DCs from 7 images.