FGFR3 activation as a function of ligand concentration, on a relative but internally consistent scale.

<p>Data shown are averages and standard errors from 3 independent experiments (one of them is presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056521#pone-0056521-g001" target="_blank">Figure 1</a>). HEK293 T cells were transfected with plasmids encoding FGFR3/WT or FGFR3/A391E. They were starved for 24 hours in serum-free medium, before <i>fgf1</i> (Millipore, MA) was added at concentrations ranging from 5 ng/ml to 5000 ng/ml (0.3 to 316 nM). After a 10 minute incubation with ligand, the cells were lysed and analyzed by Western blotting. The activation was determined by taking the ratio of anti-Y653/4 to anti-FGFR3 staining intensities in Western blot experiments such as the one shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056521#pone-0056521-g001" target="_blank">Figure 1</a>. The wild-type activation is assigned a value of 1 at saturating <i>fgf1</i> concentration <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056521#pone.0056521-Chen2" target="_blank">[20]</a>. Note that this assignment is arbitrary, and the y-axis reports on the relative, and not absolute, activation.