Figure 6.

ADV induced cell cycle arrest in G₀/G₁ phase by up-regulating p21. Resting human T cells were freshly stimulated with anti-CD3/CD28 antibodies and cultured with increasing concentrations of ADV for 72 h. DNA content and cell cycle phase of cells was determined using propidium iodide staining and flow cytometric analysis. Representative histograms of DNA content are shown in (A) (G₀/G₁ (black), S (light grey), G₂/M (dark grey). (B) The bars indicate the percentage of cells in G₀/G₁ phase, S phase and G₂/M phase. Data are presented as the mean ± SEM from n = 4 independent experiments. Resting human T cells were freshly stimulated (C, D) or pre-activated for 48 h (E, F) with anti-CD3/CD28 antibodies and cultured with increasing concentrations of ADV. After additional 24 h cells were lysed and analyzed by Western blot. (C, E) Representative Western blot images of p21 expression, β-actin was used as a loading control. (D, F) Relative expression of p21 based on densitometric quantification. Quantitative data are presented as the mean ± SEM from n = 5 (D), n = 3 (F) independent experiments. Statistical analysis was performed with One-Way ANOVA and Dunnett’s Multiple Comparison Analysis Test as post hoc test. (****P ≤ 0.0001, ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05).