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. Author manuscript; available in PMC: 2009 Jul 21.
Published in final edited form as: Nat Cell Biol. 2002 Jun;4(6):439–444. doi: 10.1038/ncb798

Figure 1. Rpr stimulates IAP auto-ubiquitination and destruction.

Figure 1

a, HEK 293T cells were transfected with GST—XIAP or GST—cIAP1 and either Rpr or vector control, in the presence of zVAD-fmk. Following precipitation with glutathione—Sepharose, GST fusion protein levels were analysed by immunoblotting with an anti-GST polyclonal antibody. b, HEK 293T cells were transfected with either wild-type ubiquitin ligase or an XIAPH467A mutant and precipitated with glutathione—Sepharose. Precipitates were immunoblotted with an antibody against GST or ubiquitin. c, HEK 293T cells were transfected with GST—XIAP and either Rpr or vector control. Cells were then radiolabelled in a pulse-chase experiment. The resulting radiolabelled GST fusion proteins were analysed by autoradiography. d, Radiolabelled IAPs were incubated in crude Xenopus egg extracts, in the presence of either Rpr peptide or vehicle control. The resulting radiolabelled protein levels were analysed by autoradiography. Equal loading was verified by Coomassie blue staining of gels (data not shown).