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. 2022 Mar 1;11(3):e12198. doi: 10.1002/jev2.12198

FIGURE 3.

FIGURE 3

The cellular uptake selectivity of LEVs. (a) Representative CLSM images of Sk‐hep1 and LO2 cells incubated with DiI‐labelled Lips and LEVs. Scale bar = 50 μm. (b) Representative flow cytometry histograms of Sk‐hep1 cells and LO2 cells treated with DiI‐labelled Lips and LEVs. (c) Mean fluorescence intensities from (b). (d) Representative CLSM images of DiI‐labelled Lips and LEVs within Sk‐hep1 and LO2 cells pretreated with heparin, RGD and YIGSR. Scale bar = 50 μm. (e) Representative flow cytometry histograms and (f) mean fluorescence intensity of Sk‐hep1 cells incubated with DiI‐labelled LEVs in the presence of heparin, RGD and YIGSR. (g) Inhibitory rate of DiI‐labelled LEVs internalised into Sk‐hep1 and LO2 cells by heparin, RGD and YIGSR. (h) Expression level of HSPG and integrin in Sk‐hep1 and LO2 cells determined by western blot. (i) Representative immunofluorescence staining images of DiI‐labelled LEVs in Sk‐hep1 or LO2 cells. Nuclei were stained with DAPI (blue). HSPG and integrin were labelled with anti‐HSPG and anti‐integrin (green), respectively. Scale bar = 20 μm. (j) Fluorescence intensity of DiI‐labelled LEVs in Sk‐hep1 cells incubated with anti‐HSPG or anti‐integrin. (k) Inhibitory rate of DiI‐labelled LEVs internalisation into Sk‐hep1 cells with anti‐HSPG or anti‐integrin pretreatment. (l) Representative flow cytometry histograms of Sk‐hep1 cells and HepG2 cells treated with DiI‐labelled Lips and LEVs. (m) Mean fluorescence intensities from (l). Data are displayed as the mean ± SD (n =  3). **p < 0.01; ****p < 0.0001.