Fig. 1

DR5 knockdown increases TRAF2 polyubiquitination (a and b), which enhances AP-1 transactivation (c), a, HEK293T cells were co-transfected with the indicated plasmids carrying the indicated genes. After 42 h, the cells were lysed for IP with anti-Flag antibody and subsequent Western blotting (WB) for the indicated proteins. The experiments were done twice with identical results. b, Whole-cell protein lysates were prepared from the indicated different cell lines and then subjected to IP with TRAF2 antibody and subsequent Western blotting (WB) for different proteins as indicated. c, HEK293T cells were co-transfected with vector (V) or Flag-TRAF2 plasmid and an control (siCtrl) or DR5 siRNA (siDR5) together with an MMP1 or AP-1 luciferase reporter construct and pCH110 plasmid. After 36 h, the cells were lysed to assay luciferase activity, which was normalized to β-gal activity. Each column represents a mean ± SD of triplicate determinations from a representative experiment. The experiments were done twice with similar results