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Figure 2 | Cell Communication and Signaling

Figure 2

From: Mast cell exosomes promote lung adenocarcinoma cell proliferation – role of KIT-stem cell factor signaling

Figure 2

Uptake of human mast cell exosomes by human lung adenocarcinoma cells. Twenty microgram of the PKH67-labelled HMC-1, HEK 293 or NIH 3T3 exosomes, or a PKH67-PBS control were added per 2 ×105 A549 cells and incubated at 37°C or 4°C for 1-48 hours. The uptake of the fluorescently labelled exosomes by A549 was detected with flow cytometry (at all the time points), fluorescence microscopy (at the 12 hour time point) and confocal microscopy (at the 4 hour time point). A) Representative graph of uptake at 4 hours at 37° detected with flow cytometry. Black curve, control cells; blue curve, PKH67-PBS control; green curve, PKH67-labelled HMC-1 exosomes. B) Representative graph of uptake at 8 hours detected with flow cytometry. Black curve, control cells; red curve, PKH67-labelled HMC-1 exosomes at 4°C; green curve, PKH67-labelled HMC-1 exosomes at 37°C. C and D) Percent positive cells and relative fluorescence intensity (rFI) data for all time points determined with flow cytometry are shown as mean × SEM (n =3). E) Uptake of PKH67-PBS control and PKH67-labelled exosomes at 37°C imaging with fluorescence microscopy. Nuclei were stained with 7-AAD (red). F) Uptake of PKH67-labelled exosomes at 37°C imaging with confocal microscopy (bar is indicating 20 μm). Nuclei were stained with DAPI (blue). G) Uptake of PKH67-labelled NIH 3T3- and HEK 293-derived exosomes at 37°C imaging with fluorescence microscopy. Nuclei were stained with 7-AAD (red).

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