Fig 4.

Virus penetration in dystonin-depleted and control cells. (A) shControl and shDyst HFFF2 cells were incubated with vSR27-VP26GFP virions for 1 h at 4°C or for 2 h at 37°C. The cells were fixed and labeled for gD with MAb 4846 and GAM₅₆₈ (red). Capsids were visualized through GFP fluorescence (green). Nuclei were visualized with DAPI (blue). Bars, 20 μm. (B) The total numbers of capsids present on randomly chosen cells from the experiment whose results are shown in panel A were counted, and the proportion having envelopes was determined by colocalization between GFP (capsid) and Alexa Fluor 568 (envelope) signals. Results are shown as the numbers of gD-positive (yellow) and gD-negative (green) capsids expressed as a percentage of the total number of capsids counted. (C) Average number of cytosolic (gD-negative) capsids per cell at 37°C. A total of 270 (ShControl, 4°C), 1,870 (shControl, 37°C), and 1,836 (shDyst, 37°C) particles were analyzed in 27, 43, and 46 cells, respectively.