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. 2016 Apr 6;12(4):671–688. doi: 10.1080/15548627.2016.1149659

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Figure 5. — Ego4 protein is identified as a negative meiotic regulator. (A-D) Free GFP accumulation after 6 h sporulation in EGO4-GFP (A), RPL37A-GFP (B), YEF3-GFP (C) and ALD6-GFP (D) WT strains but not atg12Δ strains. WT and atg12Δ strains were sporulated, and samples at 0 h and 6 h sporulation were collected for protein extraction and western blotting analysis. (E, G, H) EGO4, but not RPL37A or YEF3 overexpression inhibited yeast sporulation. EGO4 (E), RPL37A and YEF3 (G) were overexpressed under the control of the GAL1 promoter, and sporulation was induced by 0.5% galactose. Cells with empty vectors (EV) were used as a control. Samples collected at different time points were fixed for checking their sporulation rates under microscopy. The inhibitory effect of Ego4 on meiosis was compared with overexpression of a known negative meiotic regulator, Rme1 (H). (F) Ego4 overexpression delayed premeiotic DNA replication. Premeiotic DNA replications were detected by flow cytometry at different time points when Ego4 was overexpressed.