Figure 6.

Rck-mediated host cell alteration is a consequence of a DNA damage checkpoint response. HCT116 cells were infected with MC-RckGFP strain for 1 h with a MOI of 100. (A) Representative images of immunoblotting analysis corresponding to GAPDH as a protein loading control and p21 expression level of sorted GFP− and GFP+ infected cells at 3 and 6 h p.i. (B) DNA histograms generated at 0 and 6 h p.i. of sorted GFP and GFP+ infected cells maintained in the presence of caffeine for 6 h. Results are means ± SD from three independent experiments. Data were compared using a Mann-Whitney test (*p < 0.05; Untreated vs. Caffeine). (C) HCT116 cells were treated with RO-3306 for 24 h for G2/M synchronization, then infected with MC-RckGFP strain for 30 min at a MOI of 100. The cells were maintained in the presence or absence of caffeine for 0, 6 and 24 h p.i. GFP+ and GFP− cell populations were sorted by flow cytometry and cell cycle analysis was achieved using flow cytometry. Representative histograms of DNA content for 20,000 events acquired are shown, representative of two independent experiments.