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. 2021 Dec 18;10(12):3581. doi: 10.3390/cells10123581

Figure 5.

Figure 5

Pneumolysin activates platelets and induces the release extracellular vesicles (pl-Evs). (A–H) Human platelets (diluted PRP) were stimulated with PLY (100 or 500 ng/mL, 30 min), stained with CD62P (PerCP) antibody or annexin V (APC), and then analyzed by flow cytometry. (A) Percentage of CD62P positive platelets, (B) Median fluorescence intensity (MFI) of CD62P, (C) Percentage of annexin V positive platelets, and (D) Percentage of CD62P/annexin V double-positive platelets. (E) Representative FSC versus SSC dot plots of resting platelets (Ctr) or platelets treated with PLY (100 or 500 ng/mL). R1 gate includes platelets and R2 gate includes pl-EVs. (F) Quantification of the percentage of platelets and pl-EVs for each condition. (G) Representative dot plots of pl-EVs (gate R2; (E)) and (H) corresponding quantification of EVs from Ctr or PLY-treated platelets (CD41/annexin V double positive). (I) Platelet counts in whole blood of mice infected with S. pneumoniae (Spn) (48 h). * p < 0.05, ** p < 0.01, *** p < 0.001. (A–H) n = 7–9, one-way ANOVA, (F) comparisons are vs. corresponding controls, (I) n = 8–10 mice per condition, Mann–Whitney test.