Figure 6.
Association of TRAF7 with SCRIB
(A) Western blot analysis of SCRIB protein expression and its co-immunoprecipitation (co-IP) with TRAF7 using HEK293 whole-cell lysates and indicated antibodies. BAP (bacterial alkaline phosphatase), COP1, TRAF7, or TRAF7N520S were overexpressed as FLAG-fusion proteins, and co-IP was done with anti-FLAG antibody. Co-IP lanes show amount of protein in 10 times lysate input volume.
(B) Schematic structure of TRAF7 protein. S/T-rich (serine/threonine-rich), RF (ring finger), ZF (Zinc finger), CC (coiled coil), and WD40 domains are depicted by different colors. Amino acids numbers (aa #) above the diagram correspond to border amino acids in truncated FLAG-tagged TRAF7 proteins used in this study.
(C) Western blot analysis of MEKK3, MEK5, and SCRIB protein expression and their co-immunoprecipitation (co-IP) with overexpressed truncated FLAG-TRAF7 proteins using HEK293 whole-cell lysates and indicated antibodies. Co-IP was done with anti-FLAG antibody. Anti-α-Tubulin antibody was used as the loading control. Co-IP lanes show amount of protein in 10 times lysate input volume. Blots shown in panels (A) and (C) are representative of at least four independent experiments.