Figure 5.

The effect of ectopic expression of the DN TAK1 or TRAF2 mutant on Nef-induced signaling activation and macrophages colony-stimulating factor receptor (M-CSFR) downregulation in RAW264.7 cells. (a and b) RAW264.7 cells transfected with the empty vector (Empty) or the DN TAK1 or TRAF2 mutant were stimulated with Nef (100 ng/ml) for the indicated periods (min) and then analyzed by western blotting using the indicated antibodies. The data shown in the bar graphs are derived from densitometric analyses obtained from three independent experiments. (c and d) RAW264.7 cells transfected with the empty vector (Empty) or the DN TAK1 mutant were left untreated (upper panels) or stimulated with 100 ng/ml Nef (lower panels) for 2 days and then analyzed for their surface expression of M-CSFR by flow cytometry. Mean fluorescence intensity (MFI) values are shown. In the bar graph, the MFI values of the Nef-treated cells are represented as percentages relative to the MFI values of the untreated cells, and the data shown are derived from three independent experiments. *P<0.05. JNK, c-Jun N-terminal kinase