Subcellular localization of VII⁻virus particles analyzed by confocal microscopy.

A549 cells were infected with EdC-labeled (see Supplemental Information) Ad5-WT at 4^°C for 1 hour (A, surface virus) or with Ad5-WT (B, HAdV-C5_wt) or VII⁻virus particles (C, HAdV-C5_ΔVII) at 37^°C for 30 min. Cells were subsequently incubated for 0 min (30min + 0min), 3 hours (30min + 180min), or 5 hours (30min + 300min). Cells were processed for IF using antibodies directed against intact or partly fragmented Ad5 capsids (hexon, left column, white dots) followed by Click reactions to crosslink Alexa Fluor 488-azide to EdC-labeled viral DNA (center column, DNA, white dots). Panels in the right column represent merged, color images including nuclear DAPI staining. The white bar represents 10μM.