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. 2005 Mar 21;201(6):915–923. doi: 10.1084/jem.20042372

Figure 2.

Figure 2.

IRF7 activation by IRAK-1. (A) Cell lysates prepared from HEK293 cells transiently transfected with FLAG–IRF7 together with Myc–IRAK1 or Myc–IRAK1 KN were immunoprecipitated with anti-Myc or anti-FLAG, followed by immunoblot analysis using anti-FLAG or anti-Myc, as indicated. (B) HEK293 cells were transiently transfected with FLAG–IRAK-1, FLAG–IRAK-1 KN, FLAG–IRAK-4, or FLAG–IRAK-4 KN. Cell lysates were immunoprecipitated with anti-FLAG and subjected to in vitro kinase reaction in the presence of GST–IRF7. Proteins were separated on SDS-PAGE, followed by visualized by autoradiography. (C) HEK293 cells were transiently transfected with a combination of IRF7, MyD88, and 1, 10, or 50 ng of a KN mutant of IRAK-1 (IRAK-1 KN) along with a reporter plasmid carrying an IFN-α4 promoter (left). HEK293 cells were also transfected with a combination MyD88 and 1, 10, or 50 ng of IRAK-1 KN along with a reporter plasmid carrying an ELAM promoter (right). 36 h after transfection, cells were analyzed for IFN-α4– or ELAM-dependent promoter activities by a reporter gene assay.