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. 2021 Apr 7;12:650522. doi: 10.3389/fimmu.2021.650522

Figure 3.

Figure 3

Characterization and quantification of uptake of MP by unstimulated and TNFα-stimulated HUVEC. MSC were labeled with PKH-26 before generation of MP (PKH-MP). PKH-MP were added to HUVEC (ratio 1:50,000) and incubated for 4 and 24h at 37°C. (A) Uptake of PKH-MP by unstimulated and TNFα-stimulated HUVEC (ratio 1:50,000 and 1:100,000) was quantified using flow cytometry. Uptake is indicated by PKH-MP positive HUVEC (PKH+ HUVEC). (B) Representative confocal microscopy analysis of PKH-MP uptake by HUVEC at time point 24h. Staining for PKH26-MP (red), PKH-67 cell membrane (green), and Hoechst 33342 nucleus (blue) showed that PKH-MP are internalized by HUVEC. Scale bars: 40 μm (C) Staining for PKH-MP (red), lysosomes (yellow) and nucleus (blue) showed that PKH-MP (ratio 1:50,000) are co-localized with lysosomes in HUVEC after 24h of incubation. Scale bars: 20 μm.