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. 2002 Apr;22(8):2586–2597. doi: 10.1128/MCB.22.8.2586-2597.2002

FIG. 2.

FIG. 2.

Binding of CR-1 to ALK4 expressed on the cell surface of mammalian epithelial cells. (A) 293 cells transiently transfected with ALK4-HA, ActRIIB-HA, or vector alone were incubated in the presence of CR-1ΔC-Fc recombinant protein or LTβR-Fc control protein. Binding of CR-1ΔC-Fc or LTβR-Fc protein to 293 cells was analyzed by FACS analysis as described in Materials and Methods. (B) 293 cells transiently transfected with ALK4-HA or ActRIIB-HA expression vector were immunoprecipitated (IP) with CR-1ΔC-Fc or LTβR-Fc and analyzed by Western blot analysis using an anti-HA rabbit polyclonal antibody. (C) As control, cell lysates from ALK4-HA- and ActRIIB-HA-transfected 293 cells were analyzed by Western blot analysis using an anti-HA rabbit polyclonal antibody. (D) COS1 cells were transiently transfected with ALK4-Flag and/or CR-1 expression vectors and Flag-tagged proteins were immunoprecipitated with an anti-Flag mouse monoclonal antibody. The immunoprecipitated proteins were analyzed by Western blot (WB) with an anti-CR-1 rabbit polyclonal antibody or an anti-Flag mouse monoclonal antibody. MW, molecular weight in thousands.