Journal: Scientific Reports

Article Title: Proteomic profiling of plasma extracellular vesicles identifies signatures of innate immunity, coagulation, and endothelial activation in septic patients

doi: 10.1038/s41598-025-06430-x

Figure Lengend Snippet: Characterization of plasma EVs from septic patients and health controls. The concentration of EVs ( A ) and the mean EV size ( B ) isolated from the plasma of Healthy control (Control) individuals and septic patients (Sepsis) were quantified using a nanoparticle tracking analysis (Viewsizer 3000, Horiba Scientific). Comparative analysis was conducted based on EV samples from 15 septic patients and 10 HC individuals. The bar in the middle of the dots indicates the mean value. An unpaired t-test was used for the statistical analysis (**: P ≤ 0.01, and ns: not significant). ( C ) The representative size distribution for two EV samples is presented. ( D ) Western blot of Alix (PDCD6IP) and CD9 in the EVs isolated from a group of HC individuals and septic patients. Alix: PDCD6IP (Programmed cell death 6-interacting protein), CD9: tetra-spanin membrane protein.

Article Snippet: The concentration of EVs ( A ) and the mean EV size ( B ) isolated from the plasma of Healthy control (Control) individuals and septic patients (Sepsis) were quantified using a nanoparticle tracking analysis (Viewsizer 3000, Horiba Scientific).

Techniques: Clinical Proteomics, Concentration Assay, Isolation, Control, Western Blot, Membrane

Journal: British Journal of Cancer

Article Title: CD9-positive exosomes from cancer-associated fibroblasts stimulate the migration ability of scirrhous-type gastric cancer cells

doi: 10.1038/bjc.2017.487

Figure Lengend Snippet: Effect of exosomes from fibroblasts on the migration of gastric cancer cells. ( A ) The number of 1 ng of exosomes (100 μl of 10 ng ml −1 ) was counted by Nanosight Nanoparticle Tracking Analysis (Quantum Design, Tokyo, Japan). Black line is the mean value, and the red area is SD ( n =5). The particle size of exosomes was mainly distributed at 100–200 nm, and the total number of exosomes was not significantly different between CAF and NAF. The distribution pattern of exosomes size and number was not different between CAF and NAF in a same patient. ( B ) The western blot analysis showed the expressions of CD9, CD63, and CD81 in exosomes from fibroblasts. CD9 was expressed in exosomes from CaF64 and CaF65. CD63 was not expressed on any exosomes. CD81 was expressed on exosomes from NF65. ( C ) The effect of anti-CD9 antibody to uptake of exosomes. The uptake of exosomes was abrogated by CD9 blocking antibody both in OCUM-12 cells and NUGC3 cells. ( D ) Representative images of wound-healing assay. Pictures shows the initial wound mask at 0 h (grey) and the wound mask at 24 h (orange). The number of cancer cells migrating across the wound was increased by the addition of exosomes derived from CaF64 compared with that in the control. CD9 siRNA or CD9 neutralisation antibody decreased the migration-stimulating activity of exosomes from CaF64. ( E ) Effect of CD9-positive exosomes on the migration of cancer cells. The presence of CD9-positive exosomes in the cell culture significantly increased migration of NUGC-3 cells and OCUM-12 cells. The decrease rate of migration by CD9 -siRNA was evident in NUGC-3 cells. In contrast, the decrease rate of migration by the treatment of anti-CD9 antibody was evident in OCUM-12 cells. CD9 siRNA or CD9 neutralisation antibody significantly decreased the migration-stimulating activity of exosomes from CaF64. Relative wound confluence (%) was calculated as 100 × wound closure area at each time (orange)/wound area at time 0 (grey). Data are presented as means±s.d. ( F ) the effect of exosomes from siCD9 transfected CaF. Exosomes from siCD9 transfected CaF could not increase the migration activity of NUGC-3 cells and OCUM-12 cells. A full colour version of this figure is available at the British Journal of Cancer journal online.

Article Snippet: The amount and number of exosomes from 10 μg protein was quantified by Nanosight Nanoparticle Tracking Analysis (QuantumDesign, Tokyo, Japan).

Techniques: Migration, Western Blot, Blocking Assay, Wound Healing Assay, Derivative Assay, Activity Assay, Cell Culture, Transfection

Journal: Scientific Reports

Article Title: Secretome of in vitro cultured human embryos contains extracellular vesicles that are uptaken by the maternal side

doi: 10.1038/s41598-017-05549-w

Figure Lengend Snippet: Day 3 and day 5 embryos release a heterogeneous population of EVs. ( a ) Schematic overview of the isolation protocol used to obtain EVs from embryo spent media accumulated from day 1 to day 3 (D3-EVs) or from day 3 to day 5 (D5-EVs) of development. ( b ) Fresh D3-EVs or D5-EVs isolated from six embryos were negatively stained with uranyl acetate and visualized by TEM. EVs isolated from SSS-supplemented medium (SSS-EVs) are also shown (left panel). Data shown are representative of 3 independent experiments. ( c ) Concentration defined as particle/ml (upper panel) and size distribution (lower panel) profiles were determined by NTA using 50 µl of spent media from six day 3 or day 5 embryos. Size and number of SSS-EVs are also shown. Results are expressed as mean ± SEM, n = 4 (**p = 0.001, D3-EVs vs SSS-EVs). ( d ) A representative tracking analysis plot of SSS-EVs (dilution 1:500) and of D3-EVs and D5-EVs (dilution 1:500). To better visualize peaks, a different scale was used for nanoparticles of diameter higher than 175 nm. The mode, mean value, standard deviation of size and concentration for each sample are provided. The value D50 represents the median size. Similarly, 90 percent of the distribution lies below the D90 value, and 10 percent of the population lies below the D10 value.

Article Snippet: Nanoparticle tracking analysis (NTA) was performed by an expert EV laboratory (Exosomics Siena S.p.A, Siena) in order to provide a validation of the results.

Techniques: Isolation, Staining, Concentration Assay, Standard Deviation