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Fig. 1. 14-3-3ß, 14-3-3 ${epsilon}$ , and 14-3-3 ${sigma}$ bind to CDC25B in transfected cells. HEK293 (left panels) or U2OS (right panels) cells were transfected with FLAG-tagged CDC25B together with either empty vector or one of the myc-tagged 14-3-3 subtypes as described in Materials and Methods. (Top row) Lysate. Expression of CDC25B and 14-3-3-subtypes was confirmed in cell lysates with anti-FLAG antibody against CDC25B or anti-myc antibody against 14-3-3, respectively. (Middle row) IP: CDC25B ( ${alpha}$ -FLAG Ab). CDC25B was immunoprecipitated with anti-FLAG beads followed by western blotting and detection with anti-FLAG antibody to detect CDC25B and anti-myc antibody to detect CDC25B-bound 14-3-3. (Bottom row) IP: 14-3-3 ( ${alpha}$ -myc Ab. Reciprocal immunoprecipitation; CDC25B was detected in anti-myc immunoprecipitates. Protein 14-3-3 subtypes were immunoprecipitated with anti-myc antibody; the collected 14-3-3 or 14-3-3-bound CDC25B was detected by immunoblotting. v, empty vector; ß, 14-3-3ß, ${epsilon}$ ; 14-3-3 ${epsilon}$ , ${sigma}$ ; 14-3-3 ${sigma}$ .

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