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. 2011 Jun 20;286(32):28574–28583. doi: 10.1074/jbc.M111.227942

FIGURE 6.

FIGURE 6.

Crohn disease-associated mutants of NOD2 associate with RIG-I. A, HEK293 cells were transfected with Flag-RIG-I (250 ng) and EGFP-tagged wild-type (WT), R706W, G908R, or 1007fs NOD2 (250 ng). Following transfection (∼48 h), cells were lysed, and immunoprecipitations performed with anti-Flag antibody and immunoblots performed with anti-GFP (top) or anti-Flag (middle) antibodies. In parallel, immunoblots were performed for GFP in lysates of transfected cells (bottom). B, confocal micrographs of HEK293 cells transfected with EGFP-NOD2 WT, R706W, G908R, or 1007fs NOD2 (250 ng) and DsRed-RIG-I (250 ng) 48 h following transfection. Arrows denote sites of colocalization. C, luciferase assay (expressed as fold IFNβ induction versus vector controls) from HEK293 cells transfected with vector or EGFP-WT, R706W, G908R, or 1007fs NOD2 (250 ng) and DsRed-RIG-I CARDs (200 ng) and a IFNβ-promoted luciferase construct (100 ng). Data are shown as mean ± S.D. Asterisks indicate p values < 0.05 (*) or <0.001 (**). D, immunoblots for GFP and DsRed from lysates in C.