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Interaction of ASK1 with 14-3-3 is phosphorylation-dependent. 35S-labeled ASK1 was treated with potato acid phosphatase (PAP) for the indicated times with or without phosphatase inhibitors, and then incubated with 14-3-3ζ-coated beads. After washing, 14-3-3ζ-bound ASK1 was subjected to SDS/PAGE and autoradiography (Top). Middle shows phosphatase-treated ASK1. The amount of 14-3-3ζ in the beads was revealed by Coomassie blue staining (Bottom).
