Spatiotemporal dynamics of HSV genome nuclear entry and compaction state transitions using bioorthogonal chemistry and super-resolution microscopy
(ai) Representative high magnification image of an individual cell infected with HSV-1EdC (moi 10) at 0.5 hpi. Infection was as described in Fig 5. The expanded inset shows juxtaposition of uncoated compact genomes (green) and parent capsid (red) (scale bar for main image 10 μm). An example of a nucleus containing more numerous genomes is shown in panel ii. Distributions frequencies of genome numbers for approximately 200 nuclei is shown in panel (b), representing a box and whisker plot for genome number per cell nuclei at 0.5 hpi at increasing moi. Box limits represent 2nd and 3rd quartiles with the horizontal bar in the middle showing the median and whiskers showing up the 5–95% range of the total population. Exceptional outliers (less than 5% of population) are shown as individual dots. The mean value is indicated by a ‘+’. Raw data for this summary is shown in the panel (d) below. (c) Histograms for number of genomes observed in cell nuclei at 0.5 hpi at each moi. Bin width was set at 1 genome and approximately 200 nuclei for each moi were analysed for (b) and (c). (e) Distribution of total labelled foci seen in the cytoplasm versus the nucleus at each moi.