Journal: Journal of Extracellular Biology
Article Title: Monitoring Pseudoprogression Using Circulating Small Extracellular Vesicles Expressing PD‐L1 in a Melanoma Patient Treated With Immune Checkpoint Inhibitors
doi: 10.1002/jex2.70066
Figure Lengend Snippet: Small extracellular vesicle (sEV) characterization. (A) Representative immunoblots showing expression of sEV markers (Alix, TSG101, CD63 and CD9) and PD‐L1 in plasma‐derived sEVs from a melanoma patient or lysis cell line SK‐MEL‐2 (human melanoma). Grp94 is used as negative control and actin as a loading control. (B) Representative size distribution of particles isolated from plasma of melanoma patient, obtained by nanoparticle tracking analysis (NS300) with a sample image from the video used for the tracking analysis. (C) Transmission electron microscopy images of isolated sEV. Red Scale bar = 100 nm.
Article Snippet: After transferring, membranes were blocked with 5% bovine serum albumin for 1 h and incubated overnight at 4°C with specific antibodies (1/1000): CD9 (sc‐13118, Santa Cruz Biotechnology), ALIX (NB100‐65678, Novus Bio), TSG101 (sc‐7964, Santa Cruz Biotechnology), CD63 (NBP2‐4225, BioTechne), PD‐L1 (sc‐50298, Santa Cruz Biotechnology), GRP94 (ADI‐SPA‐850, Enzo Life Sciences) and β‐actin (A3854, Sigma Aldrich).
Techniques: Western Blot, Expressing, Clinical Proteomics, Derivative Assay, Lysis, Negative Control, Control, Isolation, Transmission Assay, Electron Microscopy