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. 2018 Feb 1;29(3):363–375. doi: 10.1091/mbc.E17-08-0493

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© 2018 Stieg et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 2: — SCF^Grr1 mediates Med13 degradation following H₂O₂ stress. (A) Model of the SCF^Grr1. (B) Wild-type (RSY10) and grr1∆ cells (RSY1770) harboring Med13-HA (pKC801) were treated with 0.4 mM H₂O₂ for the time points indicated and Med13 levels were analyzed by Western blot. (C) Top panel: RSY1798 (MED13-myc::KAN) was treated with 0.4 mM H₂O₂ for the time points indicated and Med13 levels were analyzed by Western blot. Tub1 levels were used as loading controls. Bottom panel: RSY1771 (grr1∆::HIS3 MED13-myc::KAN) harboring ADH1_PRO-grr1∆L was analyzed as for RSY1798. Tub1 levels were used as loading controls. (D) Yeast two-hybrid analysis of Med13 and Grr1 derivatives. Y69a cells harboring Med13-activating domain plasmid (pKC800) and either pAS2, pAS-Grr1, or pAS2-Grr1∆L binding domain plasmids were grown on –LEU, –TRP dropout medium to select for both plasmids (left panel) and on –TRP, –LEU, –HIS –ADE (middle panel), and –TRP, –LEU, –HIS, –ADE +3-AT (right panel) to test for Med13-Grr1 interaction.