Fig. 2

S100A1 expression during cell cycle progression in Hec6 and Ishikawa cells. A Stable H6-S100A1#55 (upper) and S100A1-transiently transfected Ishikawa cells for 24 h (lower) were synchronized in the G1 phase by treatment with 50 nM rapamycin, in the early S phase by 2 μg/mL aphidicolin, or in the G2/M phase by 0.25 μg/mL nocodazole for 24 h. Western blot analysis of proteins in the cytoplasmic and the nuclear fractions. B After FLAG-tagged S100A1 was transiently transfected into Ishikawa cells, the cells were treated with 50 nM rapamycin, 2 μg/mL aphidicolin, or 0.25 μg/mL nocodazole for 24 h and then stained using anti-FLAG M2 antibody. Note the cytoplasmic and nuclear S100A1 staining, independent of the cell cycle phase