HSV-1 capsid movement in NHDFs transfected with DCTN1-siRNA or expressing EB1-DN. NHDFs transfected with DCTN1 siRNA #2 (left) or expressing dominant-negative EB1 (EB1-DN; right) were infected with HSV-1 K26GFP (green). 3 hpi, nuclei were stained with Hoechst (blue). Cells were analyzed using time-lapse microscopy on a Leica DMI6000B-AFC widefield microscope with a 100× objective using an InVivo environmental chamber at 37°C. Images were captured at 1 fps for ∼330 s. The playback speed of the video is 15 fps (i.e., ∼15× real speed). In DCTN1-depleted cells or cells expressing dominant-negative EB1, viral particles remain at the cell periphery and fail to initiate long-range transport to the nucleus.
