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. 2011 Aug 1;22(15):2741–2753. doi: 10.1091/mbc.E11-05-0426

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© 2011 Leung et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 5: — Ectopic association of Sir3 and Sir4 with category 4 genes in the absence of H3K4 methylation. DamID chromatin profiling was performed on cells expressing Sir3-Dam or Sir4-Dam fusion proteins in an otherwise WT (black bars) or Δset1 (open bars) background. The percentage of methylated DNA was determined by using real-time PCR—with primers that flank candidate methylation sites—to compare relative signals for each amplicon in digested vs. undigested DNA. The percentage of methylation by Sir3-Dam or Sir4-Dam in each strain was normalized to their respective strain expressing the unfused Dam protein. Strains YM2302, YM2312, YM2304, YM2313, YM2306, and YM2314 were used in these experiments. (A) DamID comparison of Sir3 and Sir4 association with the promoter regions of the indicated genes. (B) DamID comparison of Sir3-Dam and Sir4-Dam association with candidate methylation sites [amplicons A (−964 to −836), B (−344 to −266), and C (+541 to +642) across the CTT1 gene; CTT1 is a category 4 gene involved in the cellular response to heat. Error bars show standard errors from three independent experiments.