Stress Induces Release of Extracellular Vesicles by Trypanosoma cruzi Trypomastigotes

Figure 1

Kinetics of EV release by T. cruzi trypomastigote forms. trypomastigotes were incubated in DMEM supplemented with 5% glucose at different temperatures. EV concentrations were determined at different time points by NTA (see Material and Methods). The panels show total EV concentration (a, b) and particle size (c) in the supernatants after 2 h of incubation at different temperatures (<0.05). SEM images of trypomastigotes incubated for 2 h at 4°C (d), 26°C (e), and 37°C ((f), size  μm). Panel (g) indicates the correspondent relative fluorescence (RFU) of PrestoBlue viability reagent (mean and standard deviation, ) showing no significant (NS) differences (). EVs labeled with PKH26 (/mL and 10⁹/mL) were incubated with trypomastigotes/mL (37°C). The parasites were collected at different time points, and the median of fluorescence (h) or the percentage of labeled parasites (i) was evaluated by flow cytometry. Results are average values in triplicate measurements.

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