Fig. 1: Total exosomal protein (TEP), CD44v3 and CD3 values on exosomes isolated from plasma of HNSCC patients.

a TEP concentrations of the exosome fraction obtained from plasma of n = 17 head and neck squamous cell carcinoma (HNSCC) patients at defined time-points was measured by bicinchoninic acid (BCA) assay. Results are plotted as box-and-whisker blots representing the median value, the 25th and 75th quartiles and the range. Wilcoxon related samples tests were applied to each compare pre-OP, post-OP, end (C)RT and follow-up. Mann–Whitney test was applied to compare recurrence with follow-up. *, ** or ^## and ^### correspond to p ≤ 0.05, p ≤ 0.01 and p ≤ 0.001, respectively. Asterisks and hashtags above boxes indicate the significance compared to baseline and follow-up, respectively. (C)RT = (chemo)radiotherapy. b–d Exosomes from HNSCC patients were captured using biotinylated CD63-antibodies and stained with fluorochrome-conjugated antibodies against CD44v3 (b) and CD3 (c). Surface values as determined by on-bead flow cytometry are shown as relative fluorescence intensity (RFI) compared to an appropriate isotype control. In d, the ratio between CD44v3⁺ (Tumour-derived) and CD3⁺ (T-cell-derived) exosomes is shown. Results are plotted as box-and-whisker blots representing the median value, the 25th and 75th quartiles and the range from n = 10 (pre-OP), n = 5 (post-OP, non-recurrence, recurrence) HNSCC patients. P-values were determined by Mann–Whitney test, with * or ^#, ^##, *** corresponding to p ≤ 0.05, p ≤ 0.01 and p ≤ 0.001, respectively. Asterisks, hashtags or values above boxes indicate the significance compared to following: b asterisks pre-OP, hashtag non-recurrence and recurrence, c values post-OP, d asterisks pre-OP, hashtags recurrence.