Skip to main content
. 2015 Sep 29;18(5):667–678. doi: 10.1093/neuonc/nov243

Fig. 5.

Fig. 5.

VEGFR2 gene silencing induces cellular senescence in EGFRvIII-expressing GBM cells. (A) EGFRvIII-expressing U373MG cells were transfected with VEGFR2-specific or NTC siRNA for 18 h prior to adding BrdU (100 μM) for 12 h. BrdU-positive cells (red) were detected by immunofluorescence microscopy (representative fields shown). (B) The percentage of BrdU-positive cells is shown for cells transfected with VEGFR2-specific or NTC siRNA (mean ± SEM, n = 4, ***P < .01). (C) EGFRvIII-expressing U373MG cells were transfected with VEGFR2-specific (+) or NTC siRNA for 24 h. Immunoblot analysis was performed using the indicated antibodies. Each condition is shown in duplicate. (D) EGFRvIII-expressing U373MG cells were transfected with VEGFR2-specific or NTC siRNA. Cell death was determined using the Cell Death Detection ELISA Plus Kit (mean ± SEM, n = 6, n.s., not statistically significant). (E) EGFRvIII-expressing U373MG cells were transfected with VEGFR2-specific or NTC siRNA. Senescence was determined by SA-β-Gal staining (blue). Cells were imaged by brightfield microscopy. Representative fields are shown. (F) The number of SA-β-Gal-positive cells after transfection with VEGFR2-specific or NTC siRNA was determined using ImageJ and normalized against that observed in cultures of NTC siRNA-transfected cells (mean ± SEM, n = 4, ***P < .001).