FIG. 9.

Expression of DUSP5 causes the specific nuclear translocation of both RFP-ERK2 and endogenous ERK2. (A) NIH 3T3 cells were transfected with an expression plasmid encoding RFP-ERK2, p38-RFP, or RFP-JNK, and the subcellular distribution of RFP was determined by fluorescence microscopy. (B) NIH 3T3 cells were cotransfected with wild-type DUSP5-GFP and RFP-ERK2, p38-RFP, or RFP-JNK, and the subcellular distributions of both GFP and RFP were determined by fluorescence microscopy. (C) NIH 3T3 cells were cotransfected with either DUSP5 R53,54A-GFP and RFP-ERK2 or wild-type DUSP5-GFP and RFP-ERK2 D319N (sevenmaker mutant), and the subcellular distributions of both GFP and RFP were determined by fluorescence microscopy. (D) Cos-1 cells were transfected with Myc-tagged DUSP5, and the subcellular distributions of DUSP5 and endogenous ERK were monitored by immunofluorescence with anti-Myc and anti-ERK antibodies, respectively. Experiments were performed at least three times, and representative images are shown.