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. 2012 Oct 26;287(53):44221–44233. doi: 10.1074/jbc.M112.375360

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — Gdh3 protein is stable throughout all growth stages, whereas Gdh1 is subjected to stationary phase-specific degradation. A, immunoblot analysis of the Gdh1 and Gdh3 proteins in yeast cells at different growth stages. Yeast strains (WT, Δgdh1, Δgdh3, Δgdh1Δgdh3/YCp111, Δgdh1Δgdh3/Gdh1, Δgdh1Δgdh3/YCpGdh3) were grown in SCD, and samples were taken after 24 and 48 h of culture. Cell extracts were immunoblotted and probed with anti-Gdh1, anti-Gdh3 and anti-tubulin antibodies. B, immunoblot analysis of the FLAG-tagged Gdh1 and Gdh3 proteins in yeast cells carrying the promoter-swapped derivatives of GDH1 and GDH3 at different growth stages. Yeast strains (Δgdh1Δgdh3/YCpGdh1-FLAG, Δgdh1Δgdh3/YCpGdh3-FLAG, Δgdh1Δgdh3/YCpP_GDH1-Gdh3-FLAG, and Δgdh1Δgdh3/YCpP_GDH3-Gdh1-FLAG) were grown in SCD and samples were taken after 12, 24, 48, and 72 h of culture. Cell extracts were immunoblotted and probed with anti-FLAG and anti-tubulin antibodies.