FIG 2.

Effects of endocytosis inhibitors on CVA9 infection. (A and B) A549 cells were preincubated with inhibitors (Table 1) that were also present throughout the experiment. After virus binding, the cells were incubated for 6 h. (A) The infection percentages (cells producing high amounts of CVA9 capsid protein at 6 h p.i.) of control (Ctrl and DMSO) and inhibitor-treated cells were quantified. The experiment was performed three times, and the means are shown (± standard errors [SE]). Statistical significance was calculated with a paired-sample t test (after arcsine square root transformation). ***, P < 0.001. (B) The presence of genomic (+) and complementary (−) RNA strands was analyzed with RT-PCR. The results are averages of three parallel samples (± SE). Statistical significance was calculated with a paired-sample t test. **, P < 0.01. (C) The inhibitors were introduced to the cells at different time points postinfection, and the incubation with the inhibitors was carried out until 6 h p.i., after which the infection percentages were quantified. The experiment was performed two times, and the means are shown (± SE). (D) The Rac1 inhibitor (NSC23766) was introduced to the cells at different time points. After the 1-h treatment, the medium was replaced with fresh 1% DMEM and the incubation was carried out until 6 h p.i., after which the infection percentages were quantified. The experiment was performed two times, and the means are shown (± SE). (E) The NR-CVA9 was bound to A549 cells in the darkness. The Rac1 inhibitor (NSC23766) or DMSO (a control) were introduced to the cells, and cells were incubated in the darkness for 0, 1.5, or 6 h. At these time points, the 10-min light reactions were carried out, the drug was replaced with fresh medium, and cells were transferred back into the incubator for the remaining incubation time (6 h). The infection percentages were quantified at each time point. The experiment was performed two times, and the means are shown (± SE).