Additional file 1 of MicroRNA-371–373 cluster extracellular vesicle-based communication in testicular germ cell tumors

Supplementary Material 1: Figure S1: Graphical representation of ATRA treatment schematic used for in the NT2 cell line. Figure S2: Total number of particles/ml (A, B), and mode particle size (C, D) for all separate (T)GCT cell lines lEV and sEV NTA experiments. Data shown as mean ± SEM for 3 independent experiments. Figure S3: RNA concentration measurement for the (T)GCT cell line-derived EV populations (A) and RNA concentration normalized to the number of particles: RNA per particle ratio (B). Data shown as mean ± SEM; * - p < 0.05. Figure S4: Raw figures of the western blots performed for the cell and tissue-derived EVs. Red arrow pinpoint blots that were represented in Figure 2A. Figure S5: Raw figures of the western blots performed for the plasma-derived EVs. Red arrow pinpoint blots that were represented in Figure 2B. Figure S6: Representative bright field microscopy imaging of NT2 cells 5 and 10 days after treatment start with vehicle (A,B) and ATRA (C,D). Figure S7: Western blot for pluripotency-related factors NANOG and PAX6, and for Beta-actin in vehicle and ATRA-treated NT2 cells. Figure S8: Raw figures of the western blots performed for the ATRA-treated cells. Red arrow pinpoint blots that were represented in Figure S7. Figure S9: Secretion range (NTA particles per cell ratio) for vehicle and ATRA-treated cells, in lEV (A) and sEV (B) populations. Data shown as mean ± SEM for 5 independent experiments; * - p < 0.05. Figure S10: RNA concentration measurements in tumor tissue vs non-tumoral adjacent tissue, in lEV (A) and sEV (B) populations. Data shown as mean ± SEM for 5 independent experiments; ** - p < 0.01. Figure S11: Tissue cellular levels for miR-371a-3p (A), miR-372-3p (B), miR-373-3p (C) and let-7e (D), in non-TE TGCT, TE and adjacent testicular parenchyma tissues. Data shown on a log scale as mean ± SEM; * - p < 0.05, *** -p < 0.001. Figure S12: Spearman correlation analysis for tissue cellular and conditioned medium (CM) lEV and sEV microRNA levels (A – miR-371a-3p; B – miR-372-3p, C – miR-373-3p and D – let-7e). Figure S13: Number of particles per sample for plasma-derived EVs in TGCT patients and healthy donors, lEV (A) and sEV (B) fractions. Data shown as mean ± SEM. Figure S14: MiR-371a-3p (A), miR-372-3p (B), miR-373-3p (C) and let-7e (D) plasma circulating levels for non-TE TGCT, TE and healthy donors. Data shown as mean ± SEM; ** - p < 0.01, *** - p < 0.001. Figure S15: Spearman correlation analysis for plasma circulating and EV-secreted (lEV and sEV) microRNA levels (A – miR-371a-3p; B – miR-372-3p, C – miR-373-3p and D – let-7e).