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. 2013 Jun 14;288(30):21729–21741. doi: 10.1074/jbc.M113.463885

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — The RALA GEF RLF is in complex with JIP1 and mediates RALA-induced JNK activation. A, interaction of RLF with JIP1 is independent of ROS. FLAG-RLF was immunoprecipitated, and Western blots were analyzed for HA (JIP1) and FLAG (RLF). B, interaction of RLF with JIP1 is independent of RALA. The experiment was performed as described for A. C, RLF is in one complex with JIP1 and JNK. FLAG-RLF was immunoprecipitated with an anti-FLAG antibody and eluted from the beads with FLAG peptide. The eluate was subsequently used to immunoprecipitate HA-JIP1 with anti-JIP1 antibody. Western blots were analyzed for pJNK (Thr-183/Tyr-185), HA (JIP1 and JNK), and FLAG (RLF). Scr, Scrambled; IP, immunoprecipitation.