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. 2021 Feb 18;11:4083. doi: 10.1038/s41598-021-81697-4

Figure 4.

Figure 4

Loss of syntenin expression in primary MEFs limits retroviral transduction and exosome uptake. (a) Retroviral transduction. Retrovirus encoding LUC IRES eGFP, produced using phoenix packaging cells, was incubated with MEFs (WT and with Synt KO, shown on the left) and with MCF-7 cells (Ctrl and Synt-CRISPR, shown on the right), for 48 h. Cells expressing eGFP were quantified by flow cytometry. Bar graphs represent the percentage of cells transduced by retrovirus (i.e. expressing eGFP), relative to the percentage of WT cells that was transduced (taken as 100%). (b) Exosome uptake. MEFs, either WT or Synt-KO, were incubated with exosomes derived from MCF-7 cells expressing eGFP-syntenin (eGFP-Synt), at 37 °C for 8 h, to allow for exosome uptake, and then subjected to confocal laser scanning microscopy. Left: Representative confocal micrographs of MEFs, showing the accumulation of eGFP-Synt (green) and DAPI (blue) staining of the nuclei. Exosomes loaded with eGFP-Synt yield more puncta in WT cells than in Synt-KO cells. Right: quantification of mean eGFP fluorescence per cell.