Fig. 6. Meu14 is localized at the leading edge of the forespore membrane and is required for its extension. (A) Localization of Meu14-GFP, Spo3-HA, and Sad1 during meiosis II. meu14⁺-gfp spo3::HA diploid cells (YDO121) at metaphase II (panel i and ii) and anaphase II (panels iii-v) were analyzed by immunofluorescence microscopy. Hoechst 33342 staining is blue, Meu14-GFP is green, anti-HA staining is red, and anti-Sad1 staining is yellow. (B) Psy1, a forespore membrane component, localizes abnormally in h⁹⁰meu14psy1⁺-gfp cells (YDO150). h⁹⁰psy1⁺-gfp cells (YN68) (panel (i) and h⁹⁰meu14psy1⁺-gfp cells (panels ii-iv) harboring the integrated psy1⁺-gfp gene were induced to enter meiosis II and were analyzed by immunofluorescence microscopy. Hoechst 33342 staining is blue, anti-Sad1 staining is red, and Psy1-GFP is green. (C) Frequency of the abnormal forespore formation in h⁹⁰meu14 cells. Stained cells were classified into the following 7 classes: class I, all four forespore membranes were formed next to nuclei; class II, all four membranes were formed next to nuclei but were crushed; class III, one or two forespore membranes alone were formed normally; class IV, membranes were formed but they do not properly enclose the nucleus; class V, one or two membranes were formed but others formed next to the nuclei; class VI, all four membranes were crushed and formed aggregates near the nuclei; class VII, all four membranes were formed normally. At least 250 cells were counted. (D) Localization of Meu14-GFP in the absence of Spo3 or Spo15. h⁹⁰meu14⁺-gfp cells (YDO50; panels i and ii), h⁹⁰ meu14⁺-gfp spo3::ura4⁺ cells (YDO130; panels iii and iv), and h⁹⁰ meu14⁺-gfp spo15::ura4⁺ cells (YDO10; panels v-vii) undergoing meiosis II were analyzed by immunofluorescence microscopy. Hoechst 33342 staining is blue, Meu14-GFP is green, and anti-Sad1 staining is red. Bar, 10 µm.

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