Pro-Tumorigenic Phosphorylation of p120 Catenin in Renal and Breast Cancer
Fig 3
The pp120 antibody binds p120 around the T916 site and fails to recognize it, when it is phosphorylated.
(A) Schematic of the p120 protein, depicting the different domains, the phosphorylation sites (in red lines), the different p120 isoforms and the region used as the pp120 immunogen. (B) E-cadherin (Ecad), and pp120 western blots of the p120-mutated SW48 colon cancer cells transfected with the constructs shown (1–7) or without (8), lysed and immunoprecipitated with E-cadherin. (C) Western blot of HCT116 cell lysates using either 15D2, a monoclonal T916 (cl.1) and a polyclonal T916 antibody (pl.). Molecular weights are shown on the right. (D) Western blot of HCT116 cell lysates treated with or without λ phosphatase and blotted for the monoclonal T916 (cl.1) and 15D2. (E) p120-depleted A431 cells were transfected with either murine wild type p120 isoform 3 (mp120-3A) or a construct with the T916 site mutated to alanine (mp120-3A T916A), and blotted for the monoclonal T916 (cl.1) and 15D2 antibodies. (F) Westen blot of MDCK lysates using pp120, 15D2 or T916 (cl.1) after immunoprecipitation with the same antibodies.
