Fig. 13.
Effects of CRT0066101 on the level and activity of key regulators of the cell cycle G2/M transition in xenograft bladder carcinoma tumor explants. Western blot analysis of proteins in cellular lysates from vehicle-treated (5% dextrose) and CRT0066101-treated UMUC1 tumor explants, as described in “Materials and methods”. A total of 60 or 80 µg cell extract protein was subjected to SDS-PAGE and immunoblotted with an antibody against cyclin B1, CDK1, phospho-CDK1 (Thr161, Thr14, Tyr15), p27kip1, Gadd45α, 14-3-3σ, 14-3-3ε, phospho-Chk1 (Ser317/Ser345), pan-14-3-3, Cdc25C, phospho-Cdc25C (Ser216), Myt1, and Wee1, or with a β-actin antibody as the loading control. The levels and activities of cyclin B1, CDK1, p27kip1, Gadd45α, and 14-3-3σ/ε proteins in both vehicle-treated group and CRT0066101-treated group are shown in a, while the expression and activation of Chk1, pan-14-3-3, Cdc25C, Wee1, and Myt1 proteins in vehicle-treated and CRT0066101-treated groups are presented in b