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Figure 2


Fig. 2. Effect of the hyperphosphorylation of GFAP on the association of GFAP and 14-3-3{gamma}. (A) GFAP immunoprecipitation from GFAP transfected SW13 Cl2 cells by hyperphosphorylation and dephosphorylation treatments. The cells were treated with serum-deprived medium (lanes 2 and 4) or medium containing 100 nM Calyculin A (lanes 1 and 3). Then the cell lysate was incubated with buffer alone (lanes 1 and 2) or with CIAP (lanes 3 and 4). The immunoprecipitates were blotted with anti-GFAP or anti-14-3-3{gamma} antibody. (B) GFAP immunoprecipitation from GFAP transfected SW13 Cl2 cells by kinase treatments. The cell lysate was incubated with beads and GFAP antibody. After centrifugation, GFAP binding complex was incubated with CIAP for 30 minutes and then, was treated with cdc2 kinase (lane 2) or PKC (lane 3) in relative buffer. Then fresh cell lysate was incubated with kinase treated GFAP binding complex and the final immunoprecipitates were blotted with anti-GFAP or anti-14-3-3{gamma} antibody.