Fig. 2

HMVEC-derived EVs promote GSC tumorigenic properties and endothelial transdifferentiation potential. A. Measurement of EVs/cell released by irradiated HMVECs (10 Gy and 50 Gy) relative to sham irradiated cells, used as control (0 Gy, dashed line at value 1). Results are reported as mean ± SD (n = 3). B. FACS analysis based on green fluorescence of GSCs after HMVEC-derived EV uptake (shaded histogram = GSC control sample; open histogram = GSCs incubated with EVs). C–E. Functional in vitro assays on GSC#61, GSC#61 incubated with HMVEC-derived EVs (EVs) and GSC#61 incubated with irradiated HMVEC-derived EVs (50 Gy EVs). Growth curve C: points and range lines at each day represent mean ± SD of at least two independent experiments in triplicate. D. Migration assay: values are reported as percentage relative to GSC#61 and shown as mean ± SD from two independent experiments in triplicate. E. Colony formation assay: percent colony number values from two independent experiments in triplicate were calculated over GSC#61 and are shown as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001 vs GSC#61 without EVs (Student’s t test). F Transdifferentiation assay: cytofluorimetric evaluation of CD34 expression. Values are reported as fold change relative to GSC-derived endothelial cells (GdEC) #61. ns, not significant