Figure 4. VEGF-A expression of B16F10 cells after bevacizumab treatment The effect of bevacizumab on VEGF-A expression in B16F10 cells was determined using qPCR analysis (top) and ELISA (bottom). The amount of VEGF-A mRNA expression was measured with qPCR under normoxic (black) and hypoxic (gray) conditions after 24 h with different doses bevacizumab treatment. Expression is demonstrated in normalized fold. VEGF-A mRNA expression increased in a dose-dependent manner after treatment with bevacizumab, when cells were cultured under hypoxic conditions. An ELISA was used to measure the amount of VEGF-A protein in the supernatant of UM cell lines after hypoxic exposure for 24 h. Shown is the amount of VEGF-A protein (pg/ml). Paradoxically, the amount of produced protein is reduced in the presence of bevacizumab. AVA1=equivalent human dose: 1.25 mg/5.46 ml; AVA10=10 times the equivalent human dose: 12.5 mg/ 5.46 ml; AVA20=20 times the equivalent human dose: 25 mg/5.46 ml; CO=control group: culture medium.