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

Figure 3. The pre-meiotic S phase entry defect could be rescued by GDA1 in the gda1Δ strain.

Figure 3

(a) The expression of GDA1 could rescue the sporulation process in the gda1Δ strain. The WT strain harbored the empty vector, and the gda1Δ strains harbored either the empty vector or GDA1 under the control of its own promoter. The strains were incubated in sporulation medium for 24 hrs. Sporulation efficiency was assessed by staining with DAPI. (b) Microscopic observation of the WT strain harboring the empty vector and the gda1Δ strains harboring either the empty vector or GDA1 under the control of its own promoter after sporulation induction for 24 hrs. (c) The expression of GDA1 could rescue the pre-meiotic DNA replication defect of the gda1Δ strain. The WT strain harbored the empty vector, and the gda1Δ strains harbored either the empty vector or GDA1 under the control of its own promoter. The 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 expression of of GDA1 could rescue the stabilization of Sic1p in the gda1Δ strain during sporulation. The WT strain harbored empty vector, and the gda1Δ strains harbored either empty vector or GDA1 under the control of its own promoter. The 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. (e) The expression of Gda1p during sporulation. The WT strain expressing the GDA1-TAP allele was incubated in sporulation medium, and samples were collected at different times after sporulation induction. The expression of Gda1-TAP over time was analyzed by immunoblotting with anti-TAP antibody. Pgk1p served as a loading control. Full-length blots/gels are presented in Supplementary Figure 5.