Figure 2. Subcellular localization of LIN54. (A) GC-1 cells were infected with recombinant adenovirus expressing wild-type LIN54-EGFP. At 24 h after infection, the cells were fixed and EGFP-fused proteins were detected using fluorescent microscope. Nuclei were stained with DPAI. (B) The upper panels show examples of cytoplasmic localizations of LIN54. The lower panels are the result of observing the same field of view shown in the upper panels after leptomycin B treatment. Leptomycin B treatment (20 nM) was performed for 4 h. The phase panels show the corresponding field visualized by phase-contrast microscopy. (C) GC-1 cells were cultured in 0.1% FBS for 48 h and then infected with the recombinant adenovirus. After infection, cells were cultured in 0.1% FBS for 24 h and then LIN54-EGFP was detected. Serum-starved cells were stimulated with serum for 10 h. The scale bar is 50 μM.