International Journal of Molecular Medical Science, 2024, Vol.14, No.2, 100-105 http://medscipublisher.com/index.php/ijmms 101 Blot analysis. Panel c shows the content of major phospholipids in EVs from various body fluids, finding that semen and urine have the highest proportions of PS. Panels d-f indicate that EVs from semen and urine have significantly higher contents of PS and PE compared to those from blood. Panel g reveals that the PS+PE/PC ratios are higher in semen, saliva, and urine. Panel h uses flow cytometry to detect subpopulations of EVs exposing PS. Panel i displays the proportion of PS and TSPN positive vesicles in semen EVs. The results suggest significant fluid-specific differences in the lipid composition and PS exposure of EVs. Figure 4 investigates how extracellular vesicles (EVs) rich in phosphatidylserine (PS) interfere with the viral infection mechanisms. Panel a shows that EVs from semen and other body fluids more effectively inhibit Zika virus (ZIKV) infection compared to control liposomes. Panels b-c, using fluorescence microscopy, confirm that semen EVs effectively reduce viral attachment. Panels d-e reveal that PS-rich EVs block ZIKV infection by binding to the Axl receptor. Panels f-i present "shaving" experiments conducted after treating EVs with phospholipase D, finding that removing PS exposure significantly reduces the antiviral activity of EVs, while replenishing PS restores this capability. This study highlights the critical role of PS in the antiviral mechanisms of EVs. Figure 1 PS exposure in the viral envelope of ZIKV is essential for infection
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