Multiple Functions for ORF75c in Murid Herpesvirus-4 Infection
Figure 7
ORF75c disruption impairs the transport of incoming capsids.
A. BHK-21 cells were infected (1000 genomes/cell, 6 h) with wild-type or ORF75c−.4 MuHV-4, then stained (red) for ORF65 capsid component (mAb MG-12B8), ORF25 capsid component (mAb BH-6D3), gH/gL (mAb T2C12), ORF75b (mAb CS-4A1) or ORF75c (mAb BN-6C12). Nuclei were counter stained with DAPI (blue). B. NIH-3T3 cells or NMuMG cells were infected (1000 genomes/cell, 6 h) with wild-type or ORF75c−.4 MuHV-4, then stained (red) for ORF65 (mAb MG-12B8), TK (mAb CS-4A5), gN (mAb 3F7) or gB (mAb MG-4D11). Nuclei were counterstained with DAPI (blue). C. BHK-21 cells were infected (1000 genomes/cell, 6 h) with ORF75c+ (wild type, revertant.1 or revertant.2) or ORF75c− (ORF75c−.4, ORF75c−.7, ORF75c−.9) MuHV-4, then stained for ORF65 or gB as in B. D. NIH-3T3 cells were infected (1000 genomes/cell) with wild-type or ORF75−.4 MuHV-4 for 1, 2 or 6 h. The cells were then fixed, permeabilized and co-stained for ORF25 (BH-6D3, IgG1, red) and gN (3F7, IgG2a, green), or for ORF65 (MG-12B8, IgG2a, red) and gp70 (T3B8, IgG1, green), using isotype-specific detection. Nuclei were counterstained with DAPI (blue).