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. 2019 Sep 20;110(11):3486–3496. doi: 10.1111/cas.14187

Figure 2.

Figure 2

Bone morphogenetic protein 2 (BMP‐2) induces erythropoietin‐producing hepatocellular carcinoma receptor A6 (EPHA6) tyrosine phosphorylation. A, Immunoblot analysis of TGS‐03 cells (left) and TGS‐04 cells (right) infected with ad‐LacZ or wild‐type EPHA6 (ad‐EPHA6‐WT) with or without BMP‐2 stimulation for 24 h. Cell lysates were subjected to immunoprecipitation (IP) with anti‐FLAG antibody for EPHA6, followed by phospho‐tyrosine or FLAG immunoblotting. pTyr, phospho‐tyrosine; pSMAD1/5, phospho‐SMAD1/5; TCL, total cell lysate. B, Amino acid sequence in the conserved activation loop region of EPHA2, EPHA3, EPHA4, EPHA5, EPHA6, and EPHA8. The tyrosine residue recognized by phospho‐EPHA (pEPHA) antibody when phosphorylated is highlighted in yellow (Y779 for EPHA3 and Y925 for EPHA6). C, Immunoblots of TGS‐03 cells infected with ad‐LacZ or ad‐EPHA6‐WT and treated or untreated with BMP‐2 and 0.5 μmol/L LDN193189 for 24 h. Cell lysates were subjected to IP with FLAG antibody, followed by phospho‐EPHA or FLAG immunoblotting. D, Immunoblots of TGS‐03 cells infected with ad‐EPHA6‐WT, and treated with or without BMP‐2, DMSO control or 1 μmol/L inhibitors for 24 h as indicated. LDN193, LDN193189; Dasa, dasatinib; Fore, foretinib; pSrc, phospho‐Src. E, Phylogenetic tree of the kinase domain of the EPHA receptors