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. 2002 Apr;13(4):1427–1438. doi: 10.1091/mbc.01-10-0482

Figure 8.

Figure 8

Chromatin IP of Sir complex. Chromatin immunoprecipitation was used to investigate the ability of the Sir complex to localize to the telomeres and rDNA in the sir2 mutant strains. (A) Cross-linked chromatin was immunoprecipitated with antibodies to Sir3p. To test for localization of the Sir complex at the telomeres, the DNA pulled down was probed for telomere sequences by using PCR primers corresponding to 300 base pairs from the end of the telomere at the right arm of chromosome VI. Threefold dilutions of the immunoprecipitated DNA were used in the PCR reaction. (B) Pulled down DNA was probed for sequences corresponding to an internal nonsilenced location on chromosome XII. (C) Pulled down DNA from wild type, Δsir2, and mutant 270 was probed for more distal telomere sequences by using PCR primers that recognize sequences that are 3000 base pairs from the end of the telomere at the right arm of chromosome VI. (D) Cross-linked chromatin pulled down with anti-Sir2p was blotted to a membrane by using a slot blotting apparatus and probed for rDNA sequences.