Fig. 7.

Acidification of endosomes is not required for transduction of ALDC by AdV5. (a) ALDC or 293 cells were incubated with AdV5-GFP for 24 h in the presence of various inhibitors of intracellular acidic microenvironment or with the diluents DMSO or PBS. Trans-gene expression (GFP) was measured by flow cytometry. (b) ALDC were cultured at 37 °C with fluorescein-labelled AdV5 or dextran for 4 h in the presence of normal tissue culture media or media containing 10 mM NH₄Cl, pH 7.5. After washing fluorescein, we measured MFI by flow cytometry. (c) Relative fluorescence standard curve of fluorescein–dextran at varying pH. (d) ALDC were cultured in the presence of fluorescein-labelled AdV5 or dextran. Relative fluorescence (RFU) was measured by real-time fluorometry. Each point represents the mean of cells from four different animals tested in duplicate and error bars indicate standard error of the mean.