Figure 8.

T48 and T67 do not regulate S287 dephosphorylation. (A) Recombinant N-terminal WT GST-Cdc25 protein was incubated in interphase extract treated with or without okadaic acid for 1 h at 4°C. The samples were then washed, analyzed by SDS-PAGE, and immunoblotted with antibodies against pT48, pT67 or pT138. (B) Recombinant WT, T48V, T67V or T138V N-terminal GST-Cdc25 proteins were incubated in mitotic extract for 10 min at room temperature before being retrieved, washed, and immunoblotted with antibodies against pT48, pT67, or pT138. (C) Oocytes were injected with mRNA encoding full-length WT, S287A, T48V/T67V, or T48V/T67V/S287A FLAG-tagged Cdc25 in the presence of 200 nM leptomycin B. Oocytes were monitored for GVBD over time.