Fig. 1

Nanotrapping exosomes from tissue culture supernatants. a CD63, Alix, and Actin levels from exosomes nanotrapped by either NT80 + 82 particles (lanes 3 and 4) or by NT86 particles (Ctrl NT 86; lanes 5 and 6) from Jurkat (HTLV-1 uninfected) and HUT102 (HTLV-1 infected) tissue culture supernatants were analyzed by Western blot. MT-2 whole cell extracts and molecular weight (MW) ladder are shown (lanes 1 and 2). b CD81 ELISA signals were measured from isolated exosomes as a measure of exosome concentration. Exosomes were nanotrapped (designated by NT) from HTLV-1 negative Jurkat, and HTLV-1 infected C8166 and HUT102 cells from 1 mL of starting tissue culture supernatant. The background is designated as blank. Ultracentrifuged Jurkat exosomes (ultra Jurkat exos) were also measured. Statistical analysis was performed by unpaired t-test. ns not significant; *p-value < 0.05; **p-value < 0.01 (n − 4). c Measurement of acetylcholinesterase (AchE) activity associated with exosome populations was performed. Tissue culture supernatant was measured for AchE activity from Jurkat, C8166 (C81), MT-2, and HUT102 cells both prior to nanotrapping (blue) and after addition of NT80 + 82 and subsequent exosome isolation (orange). d Tissue culture supernatants from HTLV-1 infected cells were analyzed by specialized flow cytometry for NanoFACS. Left panel represents a representative sample prior to addition of Nanotrap^® particle (Pre-NT), middle panel represents sample with addition of Nanotrap^® particle (+NT) and right panel represents sample after Nanotrap^® particle and exosomes have been removed (Post NT). NTs are shown circled in green, while 100 nm vesicles are circled in red, and noise is circled in yellow