Figure 5.

Extracellular vesicles released by VP40-producing cells at different phases of the cell cycle. 293T, V2CL, and V2CH cells were blocked at G₀ (starvation; 0.1% fetal bovine serum DMEM), G₁/S (20 mM of hydroxyurea), and G₂/M (18-hour 20 mM of hydroxyurea pretreatment, followed by release for 1 hour and subsequent treatment with 50 ng/mL of nocodazole) for 5 days. Supernatants were harvested, filtered (0.22 μm), and analyzed by ZetaView for size (peak [mode] diameter) (A) and concentration of particles (B). Statistical analysis by Student’s 2-tailed t test compares V2CL and V2CH cell cycle-blocked groups with corresponding 293T groups (†, P < .05; ††, P < .01; †††, P < .001). Additional Student’s 2-tailed t test compares cell cycle-blocked groups with controls of their own cell type (*, P < .05; **, P < .01; ***, P < .001).