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. 2002 Jun 11;99(13):8766–8771. doi: 10.1073/pnas.132636999

Figure 6.

Figure 6

Roles of TRAF6 in signalings from various members of the TNFR superfamily expressed in hair follicles. (A) Binding of TRAF6 with various members of the TNFR family known to be expressed in hair follicles. An expression plasmid encoding FLAG-tagged TRAF6 was cotransfected into 293T cells with expression plasmids for GST-tagged cytoplasmic tails of various receptors. GST pull-down assays and Western blotting analysis of cell lysates were performed as described in Materials and Methods. (B) Requirement of TRAF6 in the XEDAR signaling. Flow cytometric analysis of expression of the CD40/XEDAR chimeric receptor in wild-type (Left) and TRAF6−/− (Right) mouse embryonic fibroblasts (Lower). Cells were stained with biotinylated anti-CD40 mAb (B-B20) and streptavidin-RED670 (solid lines) or without the Ab (dotted lines). Nuclear extracts prepared from cells untreated (−) or stimulated with anti-CD40 mAb (G28–5) were subjected to electrophoretic mobility shift assays as described in Materials and Methods (Upper). An open triangle indicates NFκB-DNA complexes and a solid triangle indicates nonspecific bands.