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. 2020 Jun 26;136(12):1381–1393. doi: 10.1182/blood.2019004746

Figure 6.

Figure 6.

Parasite-derived EVs delay the erythroid maturation and induce oxidative stress. (A) EVs in conditioned medium obtained with a culture of uninfected erythroblasts (uEbCM), infected erythroblasts (iEbCM), uninfected erythrocytes (uECM) or infected erythrocytes (iECM) were observed and quantified by flow cytometry with a GPA-PC7 labeling. EV counts in iEbCM and iECM are normalized to uEbCM and uECM, respectively. Gating strategy is shown in supplemental Figure 7. (B) The size distribution of EVs purified from a culture of infected erythrocytes was determined by NTA. (C) EVs purified from a culture of infected erythrocytes were observed by electron microscopy with a negative staining. Scale bar, 100 nm. (D) Percentage of reticulocytes in erythroblast culture at 8 days after addition of 1, 5, 10, or 20 µg/mL EVs or control medium. (E) EVs in iECM or in iECM − EV were observed and quantified by flow cytometry with GPA-PC7 labeling. Percentage of reticulocytes in erythroblasts at 8 days after addition of iECM, iECM − EV, or control medium (lower left panel). (F) Percentage of oxidized erythroblasts was evaluated by flow cytometry with DHE staining 2 days after addition of 10 µg/mL EVs, iECM, iECM − EV, or control medium. Circles indicate the number of independent experiments, and error bars show the standard error of the mean. ***P < .001, **P < .01, *P < .05. ns, nonsignificant difference.