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. Author manuscript; available in PMC: 2013 Sep 27.
Published in final edited form as: Nat Commun. 2013;4:1642. doi: 10.1038/ncomms2636

Figure 3. CRL1 complexes are stabilized in the absence of Knd1p.

Figure 3

(a) Myc epitope-tagged Pof15p was expressed from a pREP3 plasmid containing an inducible promoter in strains that harbor endogenously tagged Pof10p-Myc. The strains were constructed in two backgrounds, wildtype (WT) and Δknd1. Myc-Pof15p expression was switched on by the removal of thiamine for 14 – 24 h as indicated. CRL1 complexes were immunoprecipitated with anti-Cul1p antibodies and monitored for the levels of co-precipitated Pof10p-Myc and Myc-Pof15p. As a specificity control, the 24 h lysate was used for immunoprecipitation with rabbit IgG. The level of Pof10p bound to Cul1p was quantified and plotted in a bar chart. The bottom panel shows total lysates.

(b) Same experiment as in (a) but with Myc-Pof15p expressed at low, non-competing levels. The level of Pof15p bound to Cul1p was quantified and plotted in a bar chart.