FIG. 2.

Ad5-mediated gene delivery, but not plasmid DNA, is affected by cholesterol depletion and replacement. HeLa cell were incubated with cholesterol (Ch), mβCD, or mβCD followed by cholesterol replacement (mβCD → 0.1mM Ch). Ad5-Luc was bound in the cold, and the cells were washed as described in Materials and Methods. The results are expressed as luciferase light units (l.u.) normalized to the corresponding cell numbers obtained by crystal violet measurements of parallel samples. (A) Dose dependence of mβCD and cholesterol treatments 4 h p.i. (B) Cholesterol depletion affects Ad5-Luc gene expression at different MOIs 4 h p.i. (C) Lack of effect of cholesterol depletion on luciferase expression of plasmid DNA in cells treated with mβCD or cholesterol 8 h posttransfection (p.t.) and assayed for luciferase expression 40 h p.t. (D) Time course of Ad5-Luc gene expression in control and cholesterol-depleted cells. (E) Fiber knob and cRGD peptides inhibit Ad5-Luc gene expression in control and cholesterol-depleted cells 4 h p.i. −, not present; +, present. The error bars indicate the corresponding standard errors of the mean.