Fig. 3.

AdV5 entry and trans-gene expression in ALDC is significantly different from that in CAR-expressing 293 cells. ALDC (white circles) and 293 cells (black squares) were cultured on ice (4 °C) or at 37 °C with biotinylated AdV5-GFP for 90 min followed by washing off the inoculum. (a) Biotinylated membrane-bound virus was detected by ELISA using streptavidin-HRP. (b) Cells were transduced as before and cultured for 24 h at 37 °C; expression of the trans-gene (GFP) was detected by flow cytometry. (c) ALDC and 293 cells were cultured at 37 °C in the presence of AdV5-GFP. At 1 h intervals, aliquots were washed with PBS and the cells were allowed to recover overnight after which GFP expression was measured by flow cytometry. (d) ALDC and 293 cells were cultured at 37 °C in the presence of AdV5-GFP and without washing; GFP expression was measured by flow cytometry at 1 h intervals. Each point represents the mean of cells from six different animals tested in duplicate and error bars indicate standard error of the mean.