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Fig. 7. YFP-Cdc25B3 translocates to the cytoplasm after treatment with cycloheximide or exposure to UV-C. (A) Cycloheximide induces cytoplasmic translocation of Cdc25B. HeLa cells were microinjected with YFP-Cdc25B3, treated with 100 µg/ml cycloheximide and followed by time-lapse microscopy. The time after cycloheximide addition is indicated (in minutes). (B) Representative images of cells expressing YFP-Cdc25B3, YFP-Cdc25B3(L62A) and YFP-Cdc25B3(S323G) with or without UV-C irradiation. The different constructs were transfected into HeLa cells. Five hours after transfection, cells were irradiated with 50 J/m² UV-C and fixed 1 hour later. (C) Representative images of cells expressing YFP-Cdc25B3 after UV-C treatment and addition of kinase inhibitors or leptomycin B. YFP-Cdc25B3 was transfected into HeLa cells. Four hours after transfection, cells were treated with SB202190 (SB, 10 µM), PD98059 (MEK, 50 µM), or leptomycin B (LMB, 20 nM) for 1 hour. At that point, cells were exposed to 50 J/m² UV-C and fixed 1 hour later. (D) The cytoplasmic translocation of YFP-Cdc25B3 after UV-C treatment is abolished by a p38 inhibitor. HeLa cells were transfected with YFP-Cdc25B3 and 3 hours later SB202190 was added. Four hours after transfection cells were exposed to 50 J/m² UV-C and fixed 1 hour later. Cells were scored as in Fig. 4. The graph shows the mean of 200 cells counted in three independent experiments.

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