Spatiotemporal dynamics of HSV genome nuclear entry and compaction state transitions using bioorthogonal chemistry and super-resolution microscopy
(a) Representative images of cells infected with HSV-1EdC (moi 10). Infection was synchronised as described in Fig 5 and cells fixed at 0.5 hpi (I-IV), 1 hpi (V-VIII), or 3 hpi (IX-XII) with subsequent detection by cycloaddition and immunofluorescence for ICP4 (scale bar 10 μm). Insets from panels VIII and XII are shown magnified in (b) to illustrate a shift from ICP4 association with genome foci immediately after infection but reduced or absence of association on foci remaining at the later times. (c) 3D-SIM data of a cell nucleus infected with HSV-1EdC and fixed at 2 hpi showing residual EdC labelled infecting genomes remaining as tighter foci on the periphery of replication compartments, marked by ICP4 (red) and the absence of significant ICP4 recruitment to those remaining genome foci.