8). the uptakes of nEVsthe proof that HL60 have been not CD20 on
8). the uptakes of nEVsthe proof that HL60 have been not CD20 on their plasma membranes, Completely supported by and EVsCD20 in HL60 does not express CD20 on their plasma membranes, the uptakes of nEVs and EVsCD20 in HL60 have been statistically distinct. not statistically different. 3.8. Fluorescence Microscopy Assay of EVCD20 UptakeIn Figure 9, the fluorescence microscopy pictures of 2-Methylbenzaldehyde Cancer native (A, B, and C) and targeted (D, E, and F) EVs at five /mL just after 48 h of incubation have been reported for the three cell lines (lymphocytes: A and D; Daudi: B and E; HL60: C and F). Fluorescence pictures qualitatively confirmed cytofluorimetric evaluation. Panels A, B show a low internalization of the nEVs within the cells’ cytosolic compartment in respect to the lymphocytes (D) and Daudi (E) treated with EVsCD20 . In far more detail, the evident blue fluorescence referring to the surface functionalization of EVsCD20 underlined the efficacy from the active tropism induced by the proposed bioengineering employing the CD20 antibody. In respect for the lymphocytes and Daudi cell lines, the HL60 cells, not expressing CDMembranes 2021, 11,13 ofMembranes 2021, 11, x FOR PEER Assessment markers14 of 20 on their plasma membrane, showed no internalization enhancement deriving CD20 , and the uptake of both native and targeted EVs was almost absent from the use of EVs (panels C and F, respectively).Figure 8. The graph represents the typical along with the SE of your outcomes of targeted lymphocyte-derived Figure eight. The graph represents the typical and the SE of the results of targeted lymphocyte-derived EV uptake in lymphocytes, Daudi, and HL60 cell lines at a five /mL concentration at 24 (solid color EV uptake in lymphocytes, Daudi, and HL60 cell lines at a five /mL concentration at 2415 of 20 color (solid Membranes 2021, 11, x FOR PEER Review bars) and 48 (dashed bars) hours. The table shows the resumed three-way ANOVA analysis from the bars) and 48 (dashed bars) hours. The table shows the resumed three-way ANOVA analysis from the final results; p 0.001. Independent experiments were performed three instances. final results; p 0.001. Independent experiments were performed 3 instances.3.eight. Fluorescence Microscopy Assay of EVCD20 Uptake In Figure 9, the fluorescence microscopy images of native (A, B, and C) and targeted (D, E, and F) EVs at five /mL just after 48 h of incubation had been reported for the three cell lines (lymphocytes: A and D; Daudi: B and E; HL60: C and F). Fluorescence images qualitatively confirmed cytofluorimetric analysis. Panels A,B show a low internalization in the nEVs in the cells’ cytosolic compartment in respect for the lymphocytes (D) and Daudi (E) treated with EVsCD20. In much more detail, the evident blue fluorescence referring towards the surface functionalization of EVsCD20 underlined the efficacy with the active tropism induced by the proposed bioengineering working with the CD20 antibody. In respect for the lymphocytes and Daudi cell lines, the HL60 cells, not expressing CD20 markers on their plasma membrane, showed no internalization enhancement deriving in the use of EVsCD20, and also the uptake of each native and targeted EVs was just about absent (panels C and F, respectively).Figure 9. Representative fluorescence microscopy photos of the 3 cell lines incubated for 48 h with five /mL of native Figure 9. Representative fluorescence microscopy photos from the 3 cell lines incubated for 48 h (major panels) and anti-CD20 engineered lymphocyte-derived EVs (bottom panels). Panels A and D refer to lymphocytes, B with and C and to HL60.
