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. 2022 Aug 3;11(15):2388. doi: 10.3390/cells11152388

Figure 5.

Figure 5

Extracellular vesicles from adipocytes reprogram prostate cancer cell metabolism. (A) PC3 and DU145 cells were incubated with adipocyte-derived EVs (30 μg/mL) for 24 h. Glucose consumption was then evaluated by cytofluorimetric analysis after staining with 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-Deoxyglucose (100 μM) for 30 min. Each experiment was repeated three times. Data represent mean values ± SEM and were analyzed using a t-test. *** p < 0.001 vs. PC3 or DU145 (control). (B) Cells were incubated with adipocyte-derived EVs (30 μg/mL) for 24 h. Lactate production was then evaluated by using a lactate assay kit (according to the manufacturer’s protocol). Each experiment was repeated three times. Data represent mean values ± SEM and were analyzed using a t-test. *** p < 0.001 vs. PC3 or DU145 (control). (C) Cells were incubated with adipocyte-derived EVs (30 μg/mL) for 24 h. ATP synthesis was then evaluated by using an ATP assay kit (according to the manufacturer’s protocol). Each experiment was repeated three times. Data represent mean values ± SEM and were analyzed using a t-test. *** p < 0.001 vs. PC3 or DU145 (control). (D) After adipocyte-derived EV treatment (30 μg/mL, 24 h), Western blot analysis was performed to investigate the expression levels of p-Akt and HIF-1α in PC3 and DU145 cells. Tubulin expression was evaluated as a loading control. One representative of three experiments performed is shown.