Fig. 5.

Plasma was collected from 32 patients with cancer and 28 healthy donors. Total protein yield from each isolation was measured by the Micro BCA protein assay. (a–b) Phosphorylation of AKT and ERK1/2 was measured by western blot using equal protein amounts from 21 patients and 22 healthy donors. U87 cells overexpressing EGFRvIII were used as positive control for immunostaining. (c) Signal intensities were quantified by Image J imaging analysis software and phosphorylation was measured by calculating the relative signal intensity from phosphorylated and total protein levels. Subgroup analysis was performed with patients having high EV protein levels as shown in red. (a, d–e) To determine variability in total EV kinase levels in plasma, maximum protein amounts were loaded from a subgroup of patients with NSCLC. (f) Statistical significance was tested using the 2-tailed paired Student's t-test. ns=not significant, p≤0.05, **p≤0.01, ***p≤0.001.