Figure 2.

Recombinant p7 constructs modulate intracellular pH. HEK293 cells, transfected with p7 of the HCV genotypes 1a, 2a, 3a, and 4a, were loaded with the pH-sensitive fluorescent probe LysoSensor Green DND-189 (2 μM in HEPES buffer) for 30 min and imaged with excitation at λ = 443 nm and emission at λ = 500 nm. Amantadine and rimantadine (100 μM) in HEPES buffer were added 10 min before imaging. (A) Images show transmission overlaid with the fluorescence channel (transmission) and the fluorescence channel alone. Genotypes and treatments are indicated. Mock transfection used glycine receptor constructs in the same vector. Scale bar, 10 μm. (B) Mean of LysoSensor fluorescence intensity for each treatment. Data are the average of 20 determinations from a total of three independent cell preparations. Black columns, p7-1a; light gray, p7-2a; dark gray, p7-3a; white, p7-4a. Controls: medium gray, not transfected; dashes, mock transfected. Significance (one-way ANOVA) is indicated; ^∗p < 0.05, ^∗∗p < 0.01). To see this figure in color, go online.