Fig. 5.
The phosphorylated Src*/SD interacts with PyMT in vitro and is targeted to membrane fractions. (A) WCE (500 μg) of PyMT-Src*/SD and PyMT-EV cells were subjected to IP with HA antibodies followed by IB with PyMT antibodies. *, heavy chain. (B) WCE (500 μg) of PyMT-Src*/SD and PyMT-SrcKM/SD cells were subjected to IP with PyMT antibodies followed by IB analysis with HA, CasB and PyMT antibodies. (C) Src*/SD is targeted to the membrane fraction in PyMT-expressing cells. WCE from PyMT and TAM-R cells expressing Src*/SD and EV were fractionated into cytoplasmic (S) and membrane/pellet (P) fractions as described previously in Materials and methods and analyzed by WB with pan-Cadherin, CasB, β-actin and PyMT antibodies. (D and E) Co-localization of PyMT and Src*/SD. Confocal analysis of immunofluorescence of DAPI, PyMT and Src*/SD in PyMT-Src*/SD cells was performed as described previously in Materials and methods. Bars 20 μM. (D) Insets show higher magnification of the indicated single cell. (E) Enlarged PyMT-Src*/SD cells with high-magnification z-axis images showing co-localization of Src*/SD and PyMT.