Figure 4.

Autophosphorylation of DDR2 and Effect of Dasatinib Treatment

(A and B) ELISA (A) and immunoblot (B) results show that DDR2 phosphorylation is greater in fibroblasts with the p.Tyr740Cys or p.Leu610Pro substitutions than in control fibroblasts (Ctr 1 and Ctr 2). For ELISA (A), cells were harvested, and total phosphorylated DDR2 was measured with the DuoSet IC Phospho-DDR2 kit. For immunoblot analysis (B), cells that had been serum starved overnight were harvested and examined. The antibody used targeted phospho-Tyr740-DDR2. The band representing phospho-DDR2 is marked with a red arrow.

(C) The result of treatment with 0.1 μM dasatinib (+) in control fibroblasts (left lanes), fibroblasts from individual 3 (top right lanes), and fibroblasts from individual 1 (bottom right lanes). In fibroblasts with the p.Leu610Pro and p.Tyr740Cys substitution, the band representing phospho-DDR2 (marked with a red arrow) disappeared after dasatinib treatment. All experiments were reproduced in at least two independent experiments, and representative results are shown.

Error bars illustrate the standard deviation of replicated experiments.