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. 2009 Aug 5;284(40):27544–27556. doi: 10.1074/jbc.M109.042036

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Confirmation of the microarray results for potential Zap1-repressed genes. A, S1 nuclease protection assays were performed using RNA isolated from cells grown under the same conditions as the two sets of microarray experiments. Controls for induced genes (ZRT1), repressed genes (ADH1 and ADH3), and for equal loading (calmodulin, CMD1) were included. In addition, candidate genes selected from Table 1 were also tested. The band intensities were quantified, and the fold changes are reported. These data confirmed the microarray results for these genes. E3, experiment 3; E4, experiment 4. B, zinc dose-dependent and Zap1-dependent repression of MET3, MET14 and MET16. Wild type (DY1457) cells were grown in LZM supplemented with a range of added zinc, and mRNA levels of MET3, MET14, MET16, and CMD1 were analyzed by S1 nuclease protection assay (lanes 1–4). In addition, wild type (WT) and zap1Δ mutant cells (ZHY6) were grown in low zinc (−Zn, LZM + 1 μm ZnCl₂), and RNA was isolated and analyzed by S1 nuclease protection assay (lanes 5 and 6).