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. 2008 Feb;172(2):345–357. doi: 10.2353/ajpath.2008.070696

Figure 2.

Figure 2

Binding of Cripto-1 to Cav-1 in coimmunoprecipitation experiments in COS7 cells and in FVB/N and MCF-10A mammary epithelial cells. A: COS7 cells were transiently transfected with Cav-1-myc/His, CR-1 and c-src expression vectors either alone or in various combinations, and 48 hours after transfection the cells were lysed and proteins were immunoprecipitated with anti-myc monoclonal antibody agarose beads. The myc-tagged immunoprecipitated proteins were then analyzed by Western blot analysis using anti-CR-1 rabbit polyclonal antibody, anti-c-src mouse monoclonal antibody or anti-His mouse monoclonal antibody to detect Cav-1-myc/His fusion protein. B: COS7 cells were transiently transfected with Cav-1-myc/His, CR-1 and glypican-1-Fc expression vectors either alone or in various combinations and 48 hours after transfection the cells were lysed and protein G-agarose beads were used to immunoprecipitate glypican-1-Fc fusion protein. The bound proteins were analyzed by Western blotting with anti-CR-1 rabbit polyclonal antibody, anti-His mouse monoclonal antibody, or anti-human IgG horseradish peroxidase-linked whole antibody to detect glypican-1-Fc fusion protein. C and D: Western blot analysis for CR-1, Cav-1, c-src, and glypican-1 on cell lysates of COS7 cells transiently transfected with various expression vectors as described above. E: FVB/N primary mouse mammary epithelial cells and MCF-10A human mammary epithelial cells were lysed and immunoprecipitated with an anti-CR-1 rabbit polyclonal antibody. The immunoprecipitated proteins were then analyzed by Western blotting using an anti-Cav-1 rabbit polyclonal antibody and anti-CR-1 rabbit polyclonal antibody. IP, immunoprecipitation; WB, Western blot.