Confocal and TEM images associated with The Herpes Simplex Virus pUL16 and pUL21 Proteins Prevent Capsids from Docking at Nuclear Pore Complexes

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Folders are named with corresponding figure numbers. File types include .oib, .jpg and .tif. All can be opened with NIH Image/Fiji.

Figure 2D: The localization of HSV-2 and HSV-1 capsids in the absence of pUL16 and/or pUL21. The confocal images of Vero cells in this folder were used for quantification of the percentage of HSV-1 infected cells with capsids accumulated at the nuclear envelope. Three biological replicates with n = 198–258 infected cells examined per biological replicate. Subfolder titles correspond to the different virus strains analyzed.

Figure 3E: The colocalization of HSV-2 capsids with NPCs at late times in infection. The electron micrographs of Vero cells in this folder were used for quantification of the number of A-capsids docked at NPCs per nuclear section. n = 10–26 nuclear sections examined per condition. Subfolder titles correspond to the different virus strains analyzed.

Figures 4B and 4C: Examination of super-infection inhibition in HSV-2 WT, Δ16 and Δ21 infected cells. The confocal images of Vero cells in this folder were used for quantification of cytoplasmic mCh-VP26 capsids within mock, WT, Δ16 and Δ21 infected cells. n = 10 z-projections of infected cells per condition. Additionally, these images were used for the quantification of the percentage of mCh-VP26 capsids within the nuclear periphery of mock, WT, Δ16 and Δ21 infected cells. n = 10 z-projections of infected cells per condition. Capsids at the nuclear periphery were defined as those abutting the Hoechst 33342 signal. Subfolder titles correspond to the different virus strains analyzed.

Figure 4E: Examination of super-infection inhibition in HSV-2 WT, Δ16 and Δ21 infected cells. The confocal images of Vero cells in this folder were used for quantification of nuclear WT EdC labelled viral genomes within WT, Δ16 and Δ21 infected cells. n = 177–470 infected cells examined for fluorescent EdC puncta per biological replicate. Subfolder titles correspond to the different virus strains analyzed.

Figure 5B: The effect of prior ectopic HSV-2 pUL16 and pUL21 expression on HSV-2 and PRV capsid recruitment to nuclei and subsequent virus gene expression. The confocal images of HeLa cells in this folder were used for measuring the average distance of HSV-2 capsids from the nuclear periphery in HeLa cells that had been transfected with EGFP alone (control) or in combination with HSV-2 pUL16 and/or pUL21 expression plasmids and infected for 2h with HSV-2 186 mCh-VP26. n = 43–55 capsids were examined in each transfection condition. Subfolder titles correspond to the different expression plasmids, or combinations of plasmids, that were transfected into cells prior to infection. Within each subfolder are .oif files. These .oif files are Olympus Image File descriptors that store metadata for multidimensional z-series microscopy images. Each .oif file is accompanied by a corresponding oif.files/ folder containing associated data files such as .tif images, .roi regions of interest, .pty acquisition settings, .lut lookup tables for visualization, and .bmp preview images.

Figure 5C: The effect of prior ectopic HSV-2 pUL16 and pUL21 expression on HSV-2 and PRV capsid recruitment to nuclei and subsequent virus gene expression. he confocal images of PK15 cells in this folder were used for measuring the average distance of PRV capsids from the nuclear periphery in PK15 cells that had been transfected with EGFP alone (control) or in combination with HSV-2 pUL16 and/or pUL21 expression plasmids and infected for 2h with PRV765. n = 46–60 capsids were examined in each transfection condition. Subfolder titles correspond to the different virus strains analyzed.

Figure 6B: The colocalization of pUL16-EGFP and pUL21-EGFP with HSV-2 mCh-VP26 capsids. The confocal images of HeLa cells in this folder were used for quantifying the colocalization between EGFP signal and mCh-VP26 capsid signals compared to the average EGFP background signal taken from two points 0.5μm from the analyzed capsid. n = 40 capsids per transfection condition were examined. No significant differences were seen between any of the transfection conditions when comparing the EGFP signal associated with mCh-VP26 capsids to the EGFP control. Subfolder titles correspond to the different virus strains analyzed.

Figure 7E: The colocalization of HSV capsids with NPCs in cells treated with nocodazole. The confocal images of Vero cells in this folder were used for quantification of nuclei with capsids colocalizing with NPCs in HSV-2 infected cells treated with DMSO (vehicle control) or nocodazole at 12 hpi and fixed at 18 hpi. Three biological replicates with n = 100–140 infected cells examined per biological replicate. Subfolder titles correspond to the different virus strains analyzed in the presence and absence of nocodazole.

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