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. 2014 Feb 11;289(12):8599–8611. doi: 10.1074/jbc.M113.543116

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Pmt1-Pmt2 complexes associate with the heptameric translocon and OST. Anti-HA immunoprecipitations were carried out on digitonin-solubilized total membranes. For interacting proteins 100% of the immunoprecipitated material (IP), and 10 (A–C and F), 5 (D), and 1% (E) of the input and flow-through (FT) material were resolved on SDS-polyacrylamide gels and analyzed by Western blotting. Blots were probed with antisera (see Table 3) as indicated. From the input material, ∼95% of Pmt2 and <5% of TC and OST subunits were co-precipitated with Pmt1^HA. A–C, Pmt1^HA was immunoprecipitated from strain JHY1 expressing fully functional Alg1^myc or plasmid-encoded ^mycAlg7. D, IPs from extracts of the strains BY4741 (WT), sec71Δ, and sec72Δ expressing plasmid-encoded Pmt1^HA are shown. E, anti-HA IPs were carried out on extracts of the strains SEY6210 (WT) and JHY1 (Pmt1^HA). F, HA-tagged proteins were precipitated from strains SEY6210 (WT), JHY1 (Pmt1^HA), and MLY67 (Pmt1*^HA) expressing plasmid-encoded ^mycAlg7.