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. 2009 Oct-Dec;3(4):250–258. doi: 10.4161/pri.3.4.10388

Figure 5.

Figure 5

Immunoprecipitation of Sis1 complexes. (A) [PIN+] (NS43) cells transformed with pRS415RNQ1p plasmids expressing wild-type (WT) or the indicated mutant Rnq1 proteins were harvested, and subjected to immunoprecipitation with anti-Sis1 (S), anti-Rnq1 (R), or no (N) antibodies. Immunoprecipitated proteins were analyzed by western blotting using anti-Rnq1 (αRnq1) and anti-Sis1 (αSis1) antibodies. Lanes: W, whole cell lysate; N, no immunoprecipitating antibody; S, anti-Sis1 antibody immunoprecipitation; R, anti-Rnq1 antibody immunoprecipitation. Immunoblotting was repeatedly performed using three to five independent shuffled transformants. (B) [pin−] cells transformed with pRS415RNQ1p plasmids expressing wild-type (WT) or Rnq1-L94A proteins were harvested, and subjected to the immunoprecipitation analysis as shown in (A).