Fig. 3

SW480 and SW620-derived EV uptake pathway studies in THP-1 monocytes and M0 macrophages by flow cytometry analysis. a Fluorescence intensity of THP-1 monocytes following incubation with Syto RNA select labelled EV (10 μg/mL) in the presence of uptake inhibitors: 5 μM 5-ethyl-N-isopropyl amiloride (EIPA), 80 μM dynasore hydrate, 10 μM chlorpromazine, 20 μM nystatin and 20 μM cytochalasin D. Untreated cells were used as negative control and EV-treated cells served as positive control. The graph represents mean ± SD (n = 3). b, c Flow cytometry histograms showing fluorescence intensity of THP-1 monocytes following incubation with 10 μg/mL of Syto RNA Select labeled SW480 EVs (b) or SW620 EVs (c) in the presence or absence of the uptake inhibitors. Images are representative of 3 biological replicates. d Flow cytometry analysis of M0 macrophages following incubation with Syto RNA select labelled SW480 and SW620 EVs (10 μg/mL) in the presence of uptake inhibitors: 5 μM 5-ethyl-N-isopropyl amiloride (EIPA), 80 μM dynasore hydrate, 10 μM chlorpromazine, 20 μM nystatin and 20 μM cytochalasin D. Untreated M0 cells were used as a negative control and EV-treated M0 cells served as positive control. The graph represents mean ± SD (n = 2). e, f Flow cytometry histograms showing fluorescence intensity of M0 macrophages following incubation with 10 μg/mL of Syto RNA Select labeled SW480 EVs (e) or SW620 EVs (f) in the presence or absence of uptake inhibitors. Statistical analysis carried out with a two-way ANOVA test followed by Sidak’s post-test. *p ≤ 0.05 vs. EV-treated cells of the respective monocyte-macrophage cell subset