Fig. 3

TCR activation releases extracellular vesicles carrying CD27. Levels of sCD27 and IFN-γ- released from CD4 (A) and CD8 (B) T cells in resting or stimulated with anti-CD3 when they are pretreated or not with GW4868 for 72 h as measured by Elisa. Data represent mean ± SD of 4 independent experiments. Statistical analyses were performed using ordinary one-way ANOVA. NS = not significant, p***<0.001, p****<0.0001. (C) Immunoblot for CD27 and CD9 in the whole cell lysate (W Ctrl and W CD3) and purified exosomes (Exo Ctrl and Exo CD3) from purified CD4 T cells in resting and stimulated with anti-CD3.The same amount of protein from the exosome and whole cell lysate were loaded. Quantification of CD27 (D), sCD27 (E) and CD9 (F) as a percentage of control in the whole cell lysate and purified exosomes in resting and stimulated with anti-CD3 Ab by western blotting. The experiments were repeated 3 times independently. Data represent mean ± SD. Statistical CD27 analyses were performed using one-Anova way for multiple comparisons. ****p < 0.0001