Fig. 2.

Specific dose of bevacizumab neutralizes VEGFA secretion. a VEGFA quantification (pg/ml) at 72 h by ELISA of U87, U373, and LN229 treated with specific dose (22 μg/ml, 10 μg/ml, and 0.42 μg/ml, respectively), standard dose Bev (8.3 μg/ml), and IgG as a control. b Evaluation of the relative expression of VEGFA in treated cell lines by QRT-PCR. Ct values were normalized to GAPDH and β-actin. c The expression of VEGFA was determined via Western blot after the 72-h treatments. Actin levels were used to normalize. d Immunofluorescence staining of actin-phalloidin (green) and VEGFA (red), including merged nuclei (DAPI, blue). Scale bar, 200 μm. VEGFA signal intensity quantification per cell is shown on the bottom. Data are shown as mean ± S.D. and are representative of 2 independent experiments. P values were calculated based on the 2-tailed 2-sample t test. *P < 0.05, **P < 0.01, ***P < 0.001