Figure 3. Nuclear translocation of HMW FGF2 requires Ran GTPase activity.

A. T98G cells expressing HMW FGF2 were treated with 0.1 nM either GTPγS or GDPβs, as well as control, and subjected to immunostaining by HA antibody (green). Nuclei were stained by DAPI (blue). Scale bar 10 μm. B. T98G cells as treated in A were also subjected to fractionation. Nuclear (N) and cytosolic C. fractions were analyzed by western blot using antibodies against HA to examine the subcellular distribution of HMW FGF2 protein. GAPDH and Histone H3 were used as marker for cytosolic (C) and nuclear (N) fractions respectively. C. Whole cell lysates were extracted from T98G cells as treated in A, and subjected to immunoprecipitation with HA antibody, followed by western blot analysis using antibodies against HA and Ran.