Journal: eLife

Article Title: Framework for rapid comparison of extracellular vesicle isolation methods

doi: 10.7554/eLife.70725

Figure Lengend Snippet:

Article Snippet: The following recombinant proteins were used for CD9, CD63, CD81, and albumin: ab152262 (Abcam), TP301733 (Origene), TP317508 (Origene), and ab201876 (Abcam).

Techniques: Clinical Proteomics, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Recombinant, Isolation

Journal: Free radical biology & medicine

Article Title: Exosomal biomarkers in Down syndrome and Alzheimer's disease.

doi: 10.1016/j.freeradbiomed.2017.08.028

Figure Lengend Snippet: Fig. 1. CD81 levels were increased in those with DS (unpublished data). (A) The neuron- derived exosome isolation technology captures L1CAM-positive exosomes that have an average size diameter of 117 nm, measured with Nanosight technology. Grey shading represents ± SEM from several exosome isolations demonstrating the reproducibility of the method. (B) CD81 quantification revealed that the DS cohort had over 39% more circulating neuron-derived exosomes than age-matched controls. (C) Two-way ANOVA of younger (< 35 years) and older (> 35 years) participants in the study revealed that only having DS had a significant effect on neuronal exosome secretion levels, and no age ef- fects were observed. Error bars represent an average ± SEM. Cohort obtained from the Hamlett et al. [42] manuscript.

Article Snippet: To test our hypothesis, we isolated neuron-derived exosomes as previously described [42], and quantified CD81 by enzyme-linked immunosorbent assay (ELISA) (Cusabio, Waltham, MA) against a standard curve of recombinant CD81 (Origene Technologies, Rockville, MD).

Techniques: Derivative Assay, Isolation

Journal: Environmental pollution (Barking, Essex : 1987)

Article Title: Cellular response and extracellular vesicles characterization of human macrophages exposed to fine atmospheric particulate matter.

doi: 10.1016/j.envpol.2019.07.101

Figure Lengend Snippet: Fig. 3. Characterization of extracellular vesicles derived from THP-1 exposed to PM2.5-0.3. EVs were obtained from THP-1 exposed 6 h, 24 h or 48 h to 0 (NE), 12.5 or 50 mg/mL of PM2.5-0.3. A) Graphs representing the amounts and the mean of size of EVs measured by nanoparticle tracking analysis. Significant differences were evaluated by Student's T test (*p < 0.01); B) Transmission electronic microscopy images of EVs (bar ¼ 100 nm); C) Western-blot using CD63 and CD81 antibodies on EVs obtained from each condition.

Article Snippet: After blocking, membranes were incubated with anti-CD63 or anti-CD81 primary antibodies (Novus Biologicals, Littleton, CO, USA) at 4 C overnight, washed with PBSTween (0.05% Tween 20), and incubated with a Goat anti-Mouse IgG (HþL) secondary antibody, Horseradish Peroxidase (HRP) conjugated (Life Technologies, ThermoFisher Scientific, Illkirch, France) for 60 min. After washing with PBS-Tween, antigeneantibody reactions were detected with a Chemidoc (Biorad, Marnes la Coquette, France) using the ECL Plus Detection kit (Pierce, ThermoFisher Scientific, Illkirch, France).

Techniques: Derivative Assay, Transmission Assay, Microscopy, Western Blot

Journal: Environmental pollution (Barking, Essex : 1987)

Article Title: Cellular response and extracellular vesicles characterization of human macrophages exposed to fine atmospheric particulate matter.

doi: 10.1016/j.envpol.2019.07.101

Figure Lengend Snippet: Fig. 6. Characterization of extracellular vesicles derived from primary macrophages exposed to PM2.5-0.3. EVs were obtained from pooled supernatants of primary macrophages isolated from 6 donors and exposed 48 h to 0 (NE) or 12.5 mg/mL of PM2.5-0.3. A) Graph representing the amounts of EVs measured by nanoparticle tracking analysis. Significant differences were evaluated by Student's T test (*p < 0.01); B) Graphical representation of size distribution of EVs produced by non exposed (NE) and exposed (12.5 mg/mL) primary macrophages; C) Western-blot using CD63 and CD81 antibodies on EVs obtained from each condition; D) Transmission electronic microscopy images of EVs (bar ¼ 100 nm except when indicated).

Article Snippet: After blocking, membranes were incubated with anti-CD63 or anti-CD81 primary antibodies (Novus Biologicals, Littleton, CO, USA) at 4 C overnight, washed with PBSTween (0.05% Tween 20), and incubated with a Goat anti-Mouse IgG (HþL) secondary antibody, Horseradish Peroxidase (HRP) conjugated (Life Technologies, ThermoFisher Scientific, Illkirch, France) for 60 min. After washing with PBS-Tween, antigeneantibody reactions were detected with a Chemidoc (Biorad, Marnes la Coquette, France) using the ECL Plus Detection kit (Pierce, ThermoFisher Scientific, Illkirch, France).

Techniques: Derivative Assay, Isolation, Produced, Western Blot, Transmission Assay, Microscopy