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Figure 3


Fig. 3. FGF2 treatment does not trigger nuclear accumulation of STAT1-GFP or STAT3-YFP in RCS chondrocytes. (A,B) RCS chondrocytes were transfected with STAT1-GFP or STAT3-YFP vectors. Forty-eight hours after transfection, the cells were harvested and analyzed for STAT1 and STAT3 by WB. Note the amount of STAT fusion proteins in transfected cells (lane 2) in comparison to untransfected controls (lane 1). (C,D) RCS chondrocytes were transfected with STAT1-GFP (C) or STAT3-YFP (D), grown for 48 hours and treated with FGF2, IL6 and IFN{gamma} for the indicated times, fixed and analyzed for STAT subcellular distribution by confocal microscopy. The DAPI staining indicates the cell nucleus. Note that FGF2 did not influence the distribution of either protein in contrast to positive controls (IFN{gamma}- or IL6-treated cells) that show nuclear accumulation of all the STAT1-GFP or STAT3-YFP. Also note the incomplete IL6-mediated nuclear translocation of STAT3-YFP in cells pre-treated with FGF2 for 48 hours, suggesting an interference with canonical STAT3 signaling by the FGF2 stimulus. Scale bar: 10 µm.