Fig. 3
hsEVs activated the AKT/MTOR pathway by PTEN inhibition. A, B ADSCs were incubated with nsEVs (20 µg/mL), hsEVs (20 µg/mL), MK2206 (5 µM) and rapamycin (15 µM) as indicated for 12 h, and total proteins were collected for western blotting. The protein levels of PTEN, p-AKT, AKT, p-MTOR, MTOR, p-4EBP, 4EBP, p-p70S6K, p70S6K, HIF-1α and β-actin were determined. C, D ADSCs were harvested at the indicated times for western blotting to detect HIF-1α expression. E, F ADSCs were cultured with hsEVs (20 µg/mL) and/or MG132 (10 µM) under normoxia (21% O2) or hypoxia (1% O2) for 12 h, and then, proteins were harvested for western blotting to detect HIF-1α expression. G ADSCs were fed PBS, nsEVs (20 µg/mL), hsEVs (20 µg/mL) and hEV + PX478 (25 µM) for 12 h and then cultured with medium containing H2O2 (300 nM) for another 12 h. Cell apoptosis was measured by FCM. All data are representative of three independent experiments and are shown as the mean ± SEM. (n = 3; *P < 0.05; **P < 0.01).