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. 2015 Jul 8;5:11762. doi: 10.1038/srep11762

Figure 2. GDA1 is required for pre-meiotic S phase entry.

Figure 2

The expression of Ime1p was only slightly delayed in the gda1Δ strain during sporulation. WT or gda1Δ strains expressing the IME1-3×myc allele were incubated in sporulation medium and samples were collected at different times. The expression of Ime1p-3×myc over time was analyzed by immunoblotting with anti-Myc antibody. Pgk1p served as a loading control. Full-length blots/gels are presented in Supplementary Figure 5. (b) Real-time PCR analysis of the IME1 expression level in WT and GDA1 deletion strains. (c) The pre-meiotic DNA replication was inhibited in the gda1Δ strain during sporulation. WT or gda1Δ strains were incubated in sporulation medium and samples were collected at different times after induction. DNA content was analyzed by flow cytometry to detect pre-meiotic DNA replication (2C to 4C transition). (d) The stabilization of Sic1p in the gda1Δ strain during sporulation. WT or gda1Δ strains were incubated in sporulation medium and samples were collected at different times after induction. The expression of Sic1p over time was analyzed by immunoblotting with specific anti-Sic1 antibody. Pgk1p served as a loading control. Full-length blots/gels are presented in Supplementary Figure 5.