(A–C), Confocal microscopy of live cells (CHO-K1)
transduced by Alexa-488-labeled adenoviral vectors, used at 10,000
vp/cell and complexed with FX (8 µg/ml). (A)
HAdV5wt, 30 min pi; (B, C) HAdV5F35, 3 h pi. (i), Green
channel image; (ii), phase contrast; (iii), merge of (i) and (ii).
In (C), CHO-K1 cells were transduced by recombinant
baculoviral vector expressing RFP-tagged, late endosome marker Lamp1
protein, 24 h before incubation with HAdV5F35 vector. (i), Green
channel image; (ii), phase contrast; (iii) orange channel; (iv),
merge of (i) and (iii). (D) Live HeLa cells transduced
by recombinant baculovirus expressing RFP-tagged, early endosome
marker Rab5A protein, were incubated 24 h later with
Alexa-488-labeled HAdV5wt particles without FX, at 10,000 vp/cell
and 37°C. Picture shown was taken at 20 min after incubation
with HAdV5wt. Note that most of the virus signal is weak and
diffuse, but some green fluorescent dots are visible within the
cytoplasm (white arrows). (E), Live CHO-CD46 cells
transduced by recombinant baculovirus expressing RFP-tagged, late
endosome marker Lamp1 protein, were incubated 24 h later with
Cy5-labeled HAdV3 penton dodecahedrons (Pt-Dd) at 37°C. Picture
shown was taken at 60 min after incubation with Pt-Dd. (i), Cy5
channel; (ii), phase contrast image; (iii) RFP channel; (iv), merge
of (i) and (iii).