FIG. 5.

Analyses of Ad5LucFF/6H binding to CAR. (A) ELISA employing recombinant human sCAR protein. Baculovirus-expressed sCAR protein adsorbed on an ELISA plate was incubated with various amounts of purified Ad virions. Virions bound to sCAR were then detected with anti-Ad2 polyclonal antibody. Each point represents a mean of three readings obtained in one experiment, with the error bars showing standard deviations. The background, which was equal to 0.06, has been subtracted from all data points. (B) Ad5 fiber knob-mediated inhibition of 293/6H cell transduction. 293/6H cells expressing AR and grown in the wells of a 24-well plate were preincubated with PBS or E. coli-expressed recombinant Ad5 fiber knob protein at concentrations of 1 and 10 μg/ml prior to infection with the Ad vectors. Luciferase activities detected in the lysates of cells infected in the presence of the knob are shown as percentages of the activity registered in cells in which infection was blocked with the inhibitor. Each data point corresponds to an average of three measurements less the background. The error bars show the standard deviations.