Localization and function of a eukaryotic-initiation-factor-2-associated 67-kDa glycoprotein

Figure 3 Interaction of p67 and protein kinase. COS-1 cells were co-transfected with p67 and vector alone (lane 1), K296P (lane 2), binding domain (BD) (lane 3) and K296P-KD (lane 4). COS-1 cells were also co-transfected with eukaryotic translation initiation factor 2 (eIF2α) and K296P (lane 5) as a negative control. At 48 h post-transfection, the cells were labeled with ³⁵S-Met/Cys and lysed with NP-40 lysis buffer. The expressed p67 was immunoprecipitated with anti-p67 monoclonal antibody. The total cell extract (A) and immunoprecipitates (B) were resolved on SDS-PAGE and autoradiographed.