Skip to main content
Fig. 3 | Journal of Biomedical Science

Fig. 3

From: AXL phosphorylates and up-regulates TNS2 and its implications in IRS-1-associated metabolism in cancer cells

Fig. 3

Axl binds to and phosphorylates TNS2 at tyrosine 483 and regulates cancer cells proliferation. a To demonstrate the binding of TNS2 to Axl, HEK293T cells were transfected with TNS2 or Axl and lysed. TNS2 was co-immunoprecipitated (co-IP) by Myc and Axl. b To determine the effect of phosphorylation of Axl in activation of TNS2. HEK293T cells were transfected with TNS2 or Axl and lysed. TNS2 was IP by Myc, and blotted sequentially for phosphotyrosine (pTyr) and Myc. Total cell lysates were subjected to immunoblotting with related gene expression. c TNS2 is a substrate for Axl. TNS2 protein was pulled down from U2OS cells 24 h after being transfected with TNS2, followed by in vitro kinase assay. Poly(E4Y1) is an exogenous substrate for positive control. R428 is a specific Axl inhibitor. Effects of wild-type and the phosphatase-dead C231S and the phosphorylation mutant Y483F of TNS2 on the expression of various markers in Mia PaCa2 (d) and U2OS cells (e). All cells were transfected with various plasmids, respectively, and subjected to Western blot analysis 24 h later. f Effects of wild-type and the phosphatase-dead TNS2 (C231S) and the phosphorylation-dead TNS2 (Y483F) on colony formation in Mia PaCa2 cells. KD, kinase-death; Wt, wild type

Back to article page