Fig. 3

Inhibition of EVs production increases effects of bevacizumab on U87 GBM cells' viability. a NTA of LN18 GBM cells-derived EVs following treatment with GW4869 (20 μM). LN18 GBM cells were treated for 24 h with 20 μM GW4869 or DMSO. CM was then collected, EVs were isolated and re-suspended in 100 μL filtered sterile PBS. EVs suspension was 1/5 diluted and infused to a Nanosight© NS300 instrument. 5 captures of 60s each were recorded. Particles concentration (particles/mL) and size (nm) were measured. Particles concentration was normalised to the number of cells after treatment (particles/mL/cell). b LN18 and U87 GBM cells' viability assay in response to bevacizumab combined with GW4869. Cells were seeded in a 96well plate and allowed to grow for 24 h. Cells were then treated with different bevacizumab concentrations (0.25 mg/mL or 1.5 mg/mL) with or without GW4869 (10 μM or 20 μM) for 24 h and 48 h. CellTiter-Glo® luminescent cell viability assay was then performed. Results are expressed as normalised Relative Light Unit (RLU). The mean ± SEM of 4 independent experiments is shown. (*p < 0.05, ***p < 0.01; ANOVA). c LN18 and U87 GBM cells' invasiveness assay using a hyaluronic acid (HA) hydrogel. Cells were incubated within a HA hydrogel for 7 days. Cells were treated with bevacizumab (0.25 mg/mL) with or without GW4869 (20 μM). 5 pictures (capture) per gel were taken following the treatments. Colony counting followed by CellTiter-Glo® Luminescent Cell Viability Assay were then performed. Results are expressed as normalised number of colonies / capture and normalised RLU, respectively. The mean ± SEM of 3 independent experiments is shown (*p < 0.05; ANOVA)