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. 2001 Apr 15;15(8):955–967. doi: 10.1101/gad.873601

Figure 1.

Figure 1

Sum1-1p associated with HM loci. (A) Lysates were prepared from formaldehyde cross-linked MATα haploids cells of the genotypes sir2Δ 7myc–SUM1-1 (lanes 1–4,6; JRY7153) and sir2Δ SUM1-1 (lane 5; JRY7152). DNA immunoprecipitated with anti-myc antibodies was analyzed by simultaneous PCR amplification of the HMR-E silencer region (top) and the SSC1 promoter (bottom). A total of 1/28,000 (lane 1) or 1/56,000 (lane 2) of the input DNA or 1/38 (lanes 3,5,6) or 1/76 (lane 4) of the immunoprecipitated DNA was analyzed. Negative control lanes had samples prepared with an untagged version of Sum1-1p (lane 5) or without antibody in the immunoprecipitation (lane 6). DNA molecular weight marker is 1-kB DNA ladder (GIBCO BRL). (B,C) Chromatin immunoprecipitation was performed as above using MATα haploids of the genotypes sir2Δ 7myc–SUM1-1 (lanes i, +; JRY7153) and sir2Δ SUM1-1 (lanes i; JRY7152) strains. A total of 1/28,000 input DNA (lanes i) or 1/50 of the immunoprecipitated DNA (lanes + and −) was amplified using primers to the indicated regions of HMR (B, top) or the indicated primer pair (C, top) and to the SSC1 promoter region (bottom). Negative control lanes had samples prepared with an untagged version of Sum1-1p (lanes −).