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. 2007 Jan 8;27(6):2324–2342. doi: 10.1128/MCB.02300-06

FIG. 4.

FIG. 4.

G3BP-1 binds Caprin-1 through the NTF-2-like domain. (A) Conservation of the NTF-2-like domain and RNA-binding domain of G3BP-1 in human, Xenopus, and Drosophila cells. (B) G3BP-1 mutants used. (C) The RNA-binding domain of G3BP-1 is not necessary for its interaction with Caprin-1. 293T cells were transfected with plasmids expressing Caprin-1-HA and Flag-G3BP-1 or fragment of G3BP-1 (1 to 340), and the cell lysates were subjected to anti-Flag IP. The proteins on the beads were eluted and subjected to SDS-PAGE and immunoblotting with anti-GFP and anti-Flag antibodies. (D) G3BP-1 binds Caprin-1 through its amino terminus. Cell lysates from 293T cells expressing Flag-Caprin-1 were mixed with bacterially expressed GST-G3BP fragments (1 to 309 and 229 to 466) and subjected to anti-Flag IP. The proteins on the beads were eluted and subjected to SDS-PAGE and immunoblotting with anti-GFP and anti-Flag antibodies. (E) G3BP-1 recognizes Caprin-1 through its NTF-2-like domain. 293T cells were transfected with plasmids expressing Flag-Caprin-1 or Flag-Caprin-1 (47 to 380) and GFP-G3BP-1 (1 to 141), and the cell lysates were subjected to anti-Flag IP. The precipitates were subjected to SDS-PAGE and immunoblotting with anti-GFP and anti-Flag antibody.