Figure 5

p30 enhances phosphorylation of Cdc25C at S-216. A) 40 ug of nuclear (N) or cytosolic (C) fraction prepared from either p30 expressing or Mock Jurkat T-cells were loaded on 10% SDS gel and western blot analysis was performed with anti-HA to confirm p30 expression and Histone H1 western to confirm nuclear and cytosolic fractionation. B) Western blot of nuclear (N) and cytosolic (C) extracts prepared from p30 or mock Jurkat T-cells probed with anti-Cdc2 and phosphospecific Cdc2 antibody. C) Western blot of nuclear (N) and cytosolic (C) extracts prepared from p30 or mock Jurkat T-cells, probed with anti-Cdc25C or phosphospecific anti-Cdc25C. D) Western Blot analysis of nuclear or cytosolic extracts from p30 or mock Jurkat T-cells, probed with anti-Chk1 or monoclonal phosphospecific Chk1 (S-345).