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. Author manuscript; available in PMC: 2021 Jan 1.
Published in final edited form as: J Cell Biochem. 2019 Aug 12;121(1):828–839. doi: 10.1002/jcb.29328

Figure 1. Hypoxia increases the release of extracellular vesicles (EV) from pancreatic cancer cells.

Figure 1

(A) Conditioned media from pancreatic cancer cells (MiaPaCa and AsPC1) cultured under normoxia and hypoxia (moderate, 1.0% O2; extreme, 0.1% O2) for 48 h, was collected and total EV isolated by ultracentrifugation (120,000g for 2h). (B) Differential ultracentrifugation was performed to isolate size-based sub-fractions of EV (large, 2000g × 30min; moderate, 16500g × 30min; small 120,000g × 120min). (C) To examine the release kinetics, cells were incubated under normoxia or hypoxia (1.0% O2; 0.1% O2) for 0–48 h and EVs sub-fraction isolated at 12 h time intervals. In all experiments, EV pellets were suspended in equal volume of PBS and EV release estimated by measuring the protein concentration. Data is presented as mean ±S.D. of biological replicates.