Fig. 7.

Cellular location and time course of VEGF₁₆₅a-TMR binding to NLuc-VEGFR2. (a) Confocal (upper panels) (Zeiss LSM 510) or (low panels) Structured Illumination Microscopy (SIM) super resolution images of cells pre-treated with vehicle or 1 μM cediranib for 30 min at 37 °C followed by stimulation with 10 nM VEGF₁₆₅a-TMR for a further 30 min at 37 °C. Images are representative of 3 independent experiments. (b) Influence of incubation time on NanoBRET ligand-binding characteristics. Cells were pre-treated with vehicle (solid lines) or 1 μM cediranib (dotted lines) for 30 min at 37 °C, prior to stimulation with 2 nM, 10 nM or 20 nM VEGF₁₆₅a-TMR over 120 min at 37 °C. Data (mean ± S.E.M) are expressed as a percentage of the BRET ratio obtained at 120 min with 20 nM VEGF₁₆₅a-TMR from four independent experiments.