Fig 2: Purification of plasma EVs before and after acute exercise. (A) Blood samples were taken pre and post an acute exercise bout (45 min, 70% VO2max), at 2 h recovery and after 12 weeks of combined endurance and strength training. Biopsies from adipose tissue and skeletal muscle were taken at indicated time points. Plasma EVs were isolated with size exclusion columns (SEC; qEV 35 nm) or membrane affinity columns (MEM). We profiled the expression of 178 miRNAs in MEM-purified EVs with a RT-PCR-based panel pre- and post-acute exercise (n = 13). We further measured the expression of selected miRNAs in EVs isolated with MEM in a larger sample subset at all timepoints (n = 19). Lastly, we measured miRNA expression in EVs isolated with SEC pre- and post acute exercise (n = 13). (B) Nanoparticle tracking analysis (NTA) of EVs purified with MEM and SEC. (C) Immunoblotting of EV samples pre- and post-acute exercise, isolated with SEC or MEM. (D) Quantification of CD9, CD63, CD81, ApoA1, CD41, and total protein in MEM-purified EV samples, expressed as post relative to pre acute exercise. *p-value < 0.05, paired, one-sided t-test.