Figure 3.

Effects of calcyclin binding protein/Siah-1 on cell cycle regulatory proteins. Cells were synchronized in G2-M phase with 0.2 μg/mL nocodazole for 15 h and nocodazole was removed by washing; cells were then incubated in fresh medium with (+) or without (-) gastrin for the indicated times. After treatment, cellular lysates were prepared and loaded per lane. Different blots with the same samples were detected with the indicated antibodies: Cyclin E, CDK2, p27Kip1, Skp1, Skp2, and β-actin as an internal control. Gastrin treatment induced an increase in the amount of Cyclin E protein and a decrease in the level of p27Kip1 protein during the first 8 h of treatment, whereas the levels of Skp1, Skp2, and CDK2 were not affected.