FIGURE 4.
Association of NLRC4 and NAIP5 with biotinylated flagellin fragments. HEK293E cells were transfected with His6-FLAG3-TEV tagged NAIP5 and NLRC4 and BAP-tagged flagellin fragments as indicated. The BAP-tagged flagellin fragments were biotinylated in vivo by co-expressing biotin ligase from E. coli. StrepTactin beads were used for pulldown. Proteins bound to the beads were visualized by SDS-PAGE followed by silver staining (top panel). Cleared cell lysate was analyzed on Western blot probed with anti-His tag antibody (middle and bottom panel).