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

Figure 4.

Figure 4

ORC was required for Sum1-1p association with HM loci. (A) MATα haploids of the genotypes SIR SUM1 (W303), sir2Δ SUM1-1 (JRY7152), sir1Δ SUM1-1 (JRY7173), and sir1Δ sir2Δ SUM1-1 (JRY7174) were tested for their mating ability, as in Fig. 3B. (B) Chromatin immunoprecipitation was performed as in Fig. 1, using MATα haploids of the genotypes sir2Δ ORC5 7myc–SUM1-1 (JRY7153), sir2Δ orc5-1 7myc–SUM1-1 (JRY7158), and sir2Δ orc5-1 SUM1-1 (JRY7157). Immunoprecipitated DNA was analyzed by simultaneous PCR amplification of the HML-I silencer (top) and the SSC1 promoter (bottom). A total of 1/23,000 (lanes 1,3) or 1/46,000 (lanes 2,4) of the input DNA or 1/25 (lanes 5,7,9–11) or 1/50 (lanes 6,8) of the immunoprecipitated DNA was analyzed. (C) Chromatin immunoprecipitation was performed using the same strains described in B. A total of 1/25 or 1/50 of the immunoprecipitated DNA was amplified using primers to the indicated loci (top) or to SSC1 (bottom).