RTK Phosphorylation Profiling with R&D Systems RTK Array

R&D Systems™ Proteome Profiler Human Phospho-RTK Array Kit (Catalog # ARY001B) detects multiple tyrosine phosphorylated receptors in untreated and ligand-treated cell lysates. The amount of each lysate used is indicated by each cell line tested. Array signals from scanned X-ray film images were analyzed using image analysis software.

A. Treatment of human HepG2 liver cell line with insulin (1 µg/mL) stimulated insulin R (INS R) and IGF-I R tyrosine phoshorylation 15- and 7-fold, respectively. In addition, the observed basal level EGF R of tyrosine phosphorylation was increased 3-fold by insulin treatment, most likely due to transactivation by the activated insulin/IGF-I receptors (Roudabush, F.L. et al. (2000) J. Biol. Chem. 275:22583).

B. Treatment of the human glioblastoma cell line A172 with conditioned media from the human lung cell line Wi38 (secretes Gas6) for 15 minutes, stimulated the tyrosine phosphorylation of the Gas-6 receptor, Axl. Wi38 conditioned media also stimulated the tyrosine phosphorylation of HGF R but had no effect on the high basal level of EGF R tyrosine phosphorylation.

C. Treatment of the human epidermoid carcinoma cell line A431 with recombinant HGF (100ng/mL; Catalog # 294-HGN) for 5 minutes stimulated the tyrosine phosphorylation of HGF R but did not significantly increase the high basal level of EGF R and ErbB3 phosphorylation.

D. Treatment of the human breast cancer cell line MDA-MB-453 with NRG1- ß1/HRG1- ß1 (100 ng/mL; Catalog # 396-HB) for 5 minutes increased the basal level of tyrosine phosphorylation of ErbB2, ErbB3, and ErbB4 by 4-, 1.3-, and 11-fold, respectively.