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Spatiotemporal dynamics of HSV genome nuclear entry and compaction state transitions using bioorthogonal chemistry and super-resolution microscopy

Fig 7

3D-SIM analysis of genome decompaction.

3D-SIM data of HSVEdC virions adsorbed to glass coverslips as described in Fig 4. (a) Raw data was reconstructed and individual representative particles are shown as Z-projections. Quantitative analysis was carried out on approximately 800 particles via 2D-Gaussian fitting to calculate full width half maxima in each channel with numerical summary data given in the panel. (b) 3D-SIM data of a cell infected with HSV-1EdC (moi 20) and examined at 0.5 hpi. Raw data was visualised by iso-rendering in Huygens analysis software as described in materials and methods (scale bar 1 μm). Red objects denote VP5 capsids, while green objects denote EdC-labelled genomes. Blue object is nuclear DAPI staining. (c) 3D-SIM data of HSV-1EdC genomes on coverslips compared to infected cells at 0.5 hpi and 2 hpi visualised after 3D-SIM by iso-rendering in Huygens analysis software (scale bar 1 μm). Quantitative analysis of genome volume (d) and sphericity (e) is shown as box and whisker plots. Boxes show 2nd and 3rd quartiles with a horizontal bar in the middle showing the median, while whiskers show up to 5–95% of the total population. 50 genomes were analysed for each category. Unpaired two-tailed t-tests were used for statistical results (** = p<0.005, *** = p<0.0001).

Fig 7