Figure 7.

The blockade of IL-1R tI negatively impacts on leptin upregulation of VEGF/VEGFR2 expression in 4T1 cells. Effects of leptin and IL-1R tI-blocking antibodies on levels of VEGF (A, mRNA and C, protein) and VEGFR2 (B, mRNA and D, protein) in 4T1 cells. Cells were incubated with leptin (0, 1.2 nM) and anti mouse IL-1R tI antibody (0.1 μg ml⁻¹) for 24 h. Cells incubated with non-specific species-matched IgG2b served as negative controls. The VEGF and VEGFR2 mRNA levels were quantified by real-time RT–PCR and normalised to the glyceraldehyde-3-phosphatase dehydrogenase expression. The VEGF and VEGFR2 protein were determined by ELISA and western blot (WB), respectively. The VEGFR2 results from WB were analysed by densitometric analysis (imageJ software) and normalised to β-actin as a control. (a) P<0.01 when comparing basal to leptin+control Ab and (b) P<0.05 when comparing leptin+IL-1R tI to leptin+control Ab. Data (mean±s.e.) are representative of the results derived from a minimum of three independent experiments.