FIG. 1.

Ad5-mediated gene expression requires intact MTs. (A to D) A549 cells were not treated or treated with the indicated concentrations of NOC for 1 h and then infected with β-Gal-Ad5 (A to C) or Luc-Ad5 (D) for the indicated times at 37°C. Gene expression was normalized to that of drug-free infected cells (A and B) or is reported as enzymatic units per milligram of protein per minute (C) or relative light units (D) at room temperature. The reversibility of the effects of NOC was tested by washing out the drug at 180 min, and the postentry effects of NOC were assessed by adding the drug at 180 min p.i. (C). Error bars indicate standard errors of the means. (E) Toxicity of NOC for β-Gal-Ad5-infected cells (a to c) and noninfected cells (d to f) at 7 h p.i. with ethidium bromide (a, d, and g) and calcein (b, e, and h) staining and with Nomarksi (Nom) differential interference contrast imaging (c, f, and i). Note that dead cells were EthD-1 positive and calcein negative; see, e.g., methanol (MetOH, 70% for 30 min)-treated cells (g to i). NOC-treated or infected cells were all alive. Bar, 40 μm.