Distinct eIF3 complexes. (A) eIF3 complexes associated with eIF3e and eIF3m were affinity-purified by absorption of the respective pro-A-tagged proteins to IgG beads. Bound proteins were eluted by cleavage with SDS (eIF3e) or TEV protease (eIF3m). (B) The eluates described in (A) were analyzed by immunoblotting with eIF3d antibodies. The asterisk refers to cross-reactivity of the secondary antibody with Ig light chains. (C) Lysates from cells expressing Myc-tagged Csn5p, eIF3e, and eIF3m from their endogenous genomic loci were immunoprecipitated with anti-eIF3d antisera. Coprecipitated proteins were identified by immunoblotting as indicated. Total cell lysates are shown to document expression levels of the endogenously tagged proteins. (D) Protein lysate from cells expressing protein A-tagged eIF3b from the endogenous promoter was absorbed to IgG beads, and specifically retained proteins were identified by LC-MS/MS.