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. 2004 Mar;24(6):2332–2343. doi: 10.1128/MCB.24.6.2332-2343.2004

FIG. 7.

FIG. 7.

Selective inhibition of the JNK cascade by the Dok-3-SHIP-1 complex. (A) Effect of Dok-3 polypeptides on BCR-induced MAPK activation. Pools of A20 derivatives expressing the indicated polypeptides were stimulated for various periods of time with F(ab′)2 fragments of SAM IgG (3 μg/ml). After lysis, the activation of Erk-1 and Erk-2 (top blocks), JNK (third blocks from top), and p38 (fifth blocks) was assayed by immunoblotting of equivalent amounts of total cell proteins with antibodies specific for the activated forms of MAPKs. Levels of expression of MAPKs were confirmed by immunoblotting with anti-Erk (second blocks), anti-JNK (fourth blocks), and anti-p38 (sixth blocks). (B) Impact of Dok-3 polypeptides on PMA-induced activation of JNK. The experiment was conducted as detailed for panel A, except that cells were stimulated with PMA (100 nM). (C) Effect of Dok-3 polypeptides on activation of transcription factor ATF-2. Pooled cell lines were stimulated for the indicated times with F(ab′)2 fragments of SAM IgG (3 μg/ml) or PMA (100 nM; lanes P). The activation of ATF-2 was assayed by immunoblotting of equivalent quantities of cellular proteins with anti-phospho-ATF-2 (pT69/T71).