Fig 1.

Microtubule disruption before viral inoculation reduces early AAV2 transduction. At 30 to 60 min after pretreatment with DMSO or nocodazole, HeLa cells were incubated with AAV2-CMV-GFPsc (scAAV2-GFP) particles at 4°C for 40 min, and unbound virions were washed out before transferring HeLa cells to 37°C (“pulse infection”). A video of the GFP expression was taken from 3 h to 21 h after pulse infection. (A) Snapshots of the video were taken at 3, 6, 8, 12, 16, and 21 h p.i. (B) Quantification of GFP fluorescence intensity in the snapshots with Image J. (C) HeLa cells were treated with DMSO or nocodazole at 30 to 60 min before pulse infection with scAAV2-GFP at different dosages (80, 400, 2,000, and 10,000 vgs/cell). GFP expression was measured by flow cytometry at 11 h p.i., and mean fluorescence intensity (MFI) was normalized to that of DMSO treatment. The normalized ratio of the percentage of GFP-positive cells is shown in Fig. S1 in the supplemental material.