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. 2008 Jun;19(6):2476–2487. doi: 10.1091/mbc.E08-02-0118

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© 2008 by The American Society for Cell Biology

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Figure 6. — Hierarchic recruitment of Spo13, Spo2, and Spo15 to the SPB. (A) Localization of Spo13, Spo2, and Spo15. Top, localization of Spo2 and Spo15 in the spo13Δ mutant. spo13Δ cells harboring integrated spo2-GFP (YN391) or spo15-GFP (YN90) were cultured in SSL-N sporulation medium. Middle, Spo13 and Spo15 localization in spo2Δ. spo2Δ cells harboring integrated spo13-GFP (YN459) or spo15-GFP (YN389) were cultured in SSL-N sporulation medium. Bottom, Spo2 and Spo13 localization in the spo15Δ mutant. spo15Δ cells harboring integrated spo2-GFP (YN388) or spo13-GFP (YN471) were cultured in SSL-N sporulation medium. Fixed cells at different stages of meiosis were examined after staining by DAPI and GFP and with the anti-Sad1 antibody. (B) Time course of Spo15 levels during meiosis. Wild-type cells harboring integrated spo15-HA (YN77) and spo13Δ cells harboring integrated spo15-HA (YN78) were precultured overnight in SSL+N medium and then transferred to SSL-N sporulation medium. Cells entered meiosis II at 6 h. Protein extracts were prepared at intervals and subjected to Western blot analysis with the anti-HA antibody and with the anti-α-tubulin antibody as the loading control.