Fig. 5.

Competitive binding. a, b Competitive binding to Tie2 was assessed, as described in “Experimental procedures”, using BC50 concentrations of APC (3.57 nM), Ang1 (1.75 nM), and/or Ang2 (35.09 nM). c, d Similar to a and b, except APC was used in excess at a dose of 17.86 nM. a–d Graphs show binding affinity as OD (mean ± SD) from three separate experiments. The no APC control was subtracted from each data point. e Binding assays were conducted with increasing concentrations of APC and BC50 concentrations for Ang1 (full line) and Ang 2 (dashed line). Results using the stated concentration of competitor are expressed as a percentage inhibition of binding to Tie2 in the absence of competitor. Each curve represents results from three separate experiments. *P < 0.05, **P < 0.01, ***P < 0.001, n = 3. IC50 data on right was obtained from competitive binding assays and derived from three independent experiments, conducted in duplicate