Fig. 2: Impact of CD9/CD63 depletion on donor EVs using the NLuc assay.

A HeLa Cell and EV contents were assessed for CD9, CD63 and Calnexin by western blot. b) WT, CD63KO and CD9KO cells were transfected with NLuc-HSP70 reporter and EVs were isolated. B Luminescence activity was tested in EVs emanating from the three types of donor cells (equal amount of proteins). Two independent experiments were performed in duplicate, dot represent the average of each duplicate for each experiment, error bars represent the standard deviations. C EVs isolated by sequential ultracentrifugation and emanating from WT, CD63KO and CD9KO cells were analyzed by nanoparticle tracking. Graphs represent the particle size distribution for each sample. D Graph showing the EV-uptake (% from the input) in HeLa WT cells, after incubation for 24 h with EVs emanating from WT, CD63KO and CD9KO cells. Three experiments were performed, each including duplicates. Dots represent individual replicate, error bars represent the standard deviations. E Graph representing the content delivery within the cytosol of WT, when EVs emanating from WT, CD63KO and CD9KO cells were added for 24 h. Value in WT cells (control) was set to 1. Three independent experiments were performed, each dot represents an experiment. Error bars represent the standard deviations.