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. 2008 Jul 11;283(28):19245–19254. doi: 10.1074/jbc.M800943200

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Copyright © 2008, The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — TAB4 associates with and directly binds TAK1. A, FLAG-TAK1 and/or T7-TAB1 were expressed with and without HA-TAB4 in HEK293T cells and immunoprecipitated using anti-FLAG M2 beads. Extracts (lanes 1–6) and immunoprecipitates (IP)(lanes 7–12) were analyzed by immunoblotting with anti-HA, anti-T7, and anti-FLAG (top to bottom). B, empty vector, FLAG-TAK1, FLAG-TAK1 and T7-TAB1, or FLAG-TAK1, T7-TAB1, and HA-TAB4 were expressed in HEK293T cells. Proteins were immunoprecipitated using anti-FLAG M2 beads, resolved by SDS-PAGE, and analyzed by immunoblotting using anti-FLAG (upper panel). A duplicate filter was used in an overlay assay (far-Western) with 5 μg/ml GST-TAB4 or GST as probes and immunoblotted with anti-GST for detection.