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Fig. 7. PP2A, rather than PP1, is responsible for the dephosphorylation of p115HEF1 in human dermal fibroblasts. (A) The activities of PP1 and PP2A in the cell extracts obtained as described in Fig. 6 were determined by measuring the generation of free ³²P_i from ³²P-phosphorylase-a and differentiated by their sensitivities to inhibitor-2 as described in Materials and Methods. The activity of control was set as 100%. The data represent the mean ± s.d. of duplicate assays from three separate experiments. (B) To ensure equal amounts of protein phosphatases in the sample shown in A, cell extracts containing the same amount of protein were processed for western blotting with anti-PP2A monoclonal antibody. Nitrocellulose membrane was stripped and reprobed with anti-PP1 monoclonal antibody. (C) The cell extracts were obtained as described in Fig. 6. The PP1 and PP2A activities in cell extracts were determined as described above, and expressed as percentage of control. The data represent the mean ± s.d. of triplicate assays from two separate experiments. (D) Cell extracts containing the same amount of protein were analyzed by western blotting with anti-PP1 monoclonal antibody. Nitrocellulose membrane was stripped and reprobed with anti-PP2A monoclonal antibody.

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