Figure 1.

Hypoxia enhances small EV release in several cell lines. Several cell types including 3T3L1 adipocytes, NIH-3T3 fibroblasts, HEK293T, hUMSCs, AC16 and MDA-MB231were cultured and differentiated as indicated in methods section. Cells were washed in PBS and media changed to serum free or supplemented with 2–5% of exosome-free serum. Cells were exposed to hypoxia for 24 h with the exception of AC16 cells which were treated for 16 h and hUMSCs for 2h. Control cells were kept in normoxia for the same amount of time. The conditioned media was collected and small EVs were isolated by differential ultracentrifugation and quantified by NTA analysis. Data shows the mean ± standard error of the mean (SEM) of N = 3–4 independent experiments. Data were normalized to the protein content in the cellular lysates. Statistical Analysis two tailed T-test. *indicates < 0.05, **p < 0.01, ***p < 0.001. NOR normoxia, HYP hypoxia.