FIGURE 3:
Validation of the architectural model of IFT-A. (A) Interaction map of IFT-A predicted from the data shown in Figures 1 and 2. (B, C) Interaction of TULP3 with the entire IFT-A complex. HEK293T cells were cotransfected with expression vectors for EGFP-TULP3 and for all but one (IFT43 or IFT140) of the other IFT-A subunits fused to mChe, and (B) lysates prepared from the cells were processed for the VIP assay. (C) Proteins bound to the precipitated beads (lanes 4–6) or input proteins (lanes 1–3) were subjected to immunoblotting with an anti-RFP (top), anti-IFT139 (middle), or anti-GFP antibody (bottom). (D, E) Interaction of IFT139 with the other IFT-A subunits. Lysates prepared from HEK293T cells cotransfected with an expression vector for EGFP-IFT139 and for all but one (IFT43 or IFT144) of the other IFT-A subunits fused to mChe were processed for the VIP assay (D) or immunoblotting (E), as indicated.