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. 2004 Apr 8;23(9):1957–1967. doi: 10.1038/sj.emboj.7600204

Figure 2.

Figure 2

The initiation of meiotic DNA replication is delayed in set1Δ cells independently of the Clb5 function. (A) Sporulation was induced in WT CLB5myc and set1Δ CLB5myc cells. Samples were collected at the indicated times and used to analyze meiotic DNA replication (1), phosphorylation of Orc6 (2), Clb5myc accumulation (3), and kinase activity of immunoprecipitated Clb5myc (4). (1) Meiotic DNA replication was followed by FACS analysis; (2) kinetics of Orc6 phosphorylation was monitored by Western blot using anti-Orc6 antibodies (Weinreich et al, 1999); (3) Clb5myc accumulation was monitored by Western blot using anti-myc antibodies. Asterisk: degraded form of Clb5myc. Below: Ponceau red staining. (4) Kinase activity of the immunoprecipitated Clb5myc was assayed with histone H1 as a substrate. (B) Sporulation rates of the indicated strains were determined after 24, 36, and 48 h after plating on sporulation plates by measuring the ratio of sporulated versus non-sporulated diploid cells. (C) FACS analysis after induction of sporulation of clb5Δ and set1Δclb5Δ strains.