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Fig. 5. TRIP6 interacts with signaling components in the IL-1, TLR2 and Nod1 pathways. (A) TRIP6 interacts with IL-1 receptors, TLR2 and Nod1 but not MEKK3. 293 cells (2x10⁶) were transfected with 5 µg each of HA-TRIP6 and Flag-tagged IL-1Rs (IL-1R+IL-1RAcP), TLR2, Nod1 or MEKK3. Cell lysates were immunoprecipitated with anti-Flag antibody ( ${alpha}$ F) or control mouse IgG (C). The immunoprecipitates were analysed by western blots with horseradish-peroxidase (HRP)-conjugated anti-HA antibody (right). Expression of the transfected proteins in the lysates was detected by western blot analysis withanti-Flag and anti-HA antibodies (left). (B) TRIP6 interacts with MyD88, Tollip, IRAK and TRAF6. 293 cells (2x10⁶) were transfected with 5 µg each of plasmids encoding HA-TRIP6 and Flag-tagged IRAK, Tollip or TRAF6, or Myc-tagged MyD88, as indicated. Before immunoprecipitation, cells were treated with IL-1 (+) or left untreated (-), as indicated, for 2 minutes. Cell lysates were immunoprecipitated with anti-Flag antibody ( ${alpha}$ F), anti-Myc ( ${alpha}$ M) or control mouse IgG (C). The immunoprecipitates were analysed by western blots with HRP-conjugated anti-HA antibody (top). The same blots were detected again with anti-Myc or anti-Flag antibody as indicated (middle). Expression of transfected HA-TRIP6 in the lysates was detected by western blot analysis with anti-HA antibody (bottom).

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