Skip to main content
. 2003 Apr 7;100(9):5193–5198. doi: 10.1073/pnas.0531290100

Figure 2.

Figure 2

Covalent crosslinking of [125I]activin-A to type II activin receptors, Cripto, and ALK4. (A) 293T cells were transfected with the following constructs: lane 1, vector; lane 2, ActRII-myc; lane 3, ActRII-myc + ALK4; lane 4, ActRII-myc + Cripto; lane 5, ActRII-myc + Cripto mCFC; lane 6, ActRII-myc + Cripto ΔEGF; lane 7, ActRII-myc + ALK4 + Cripto; lane 8, ActRII-myc + ALK4 + Cripto mCFC; lane 9, ActRII-myc + ALK4 + Cripto ΔEGF. (B) 293T cells were transfected with the same constructs as described for A but with ActRIIB instead of ActRII-myc. (C) 293T cells were transfected with vector (lane 1), ALK4 + Cripto (lane 2), ActRII + ALK4 (lane 3), or ActRII + ALK4 + Cripto (lane 4). Cells were subjected to crosslinking with [125I]activin-A as described in Materials and Methods. Crosslinked complexes were isolated by immunoprecipitation by using an anti-myc antibody (targeting ActRII-myc) (A), an ActRIIB antibody (B), or an anti-ALK4 antibody (C). Immunoprecipitated proteins were resolved by SDS/PAGE and visualized by autoradiography as described in Materials and Methods. (D) 293T cells transfected with vector (lane 1), ActRIIB + ALK4 (lane 2), or ActRIIB + ALK4 + Cripto (lane 3) were solubilized and subjected to SDS/PAGE and Western blot analysis as described in Materials and Methods.