Time-lapse of EB1-DN in NHDFs, and HSV-1 motility in ciliobrevin D–treated NHDFs. (A) NHDFs were transfected with pN1-EB1-mCherry (false-colored green for contrast) encoding dominant-negative EB1 (EB1-DN) using Lipofectamine 3000. 24 h after transfection, cells were analyzed using time-lapse microscopy on a DMI6000B-AFC widefield microscope with a 100× objective. Images were acquired at 4 fps for 37 s. The playback speed of the video is 15 fps (i.e., ∼4× real speed). A representative cell is shown. (B) NHDFs were treated with 200-µM ciliobrevin D for 1 h and then infected with HSV-1 containing a GFP-tagged capsid protein, VP26 (K26GFP; green). 3 hpi, nuclei were stained with Hoechst (blue) and cells were analyzed using time-lapse microscopy on a DMI6000B-AFC widefield microscope with a 100× objective using an environmental chamber at 37°C. Images were captured at 1 fps for 155 s. The playback speed of the video is 15 fps (i.e., ∼15× real speed). A representative video illustrates the brief but abortive long-range motility of virus particles in the presence of dynein inhibitor.
