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Fig. 2. slk1 expression is meiosis-specific. (A) Northern-blot analysis of slk1 expression in a pat1-driven meiosis. Strain h–/h– pat1-114/pat1-114 (S964) was induced to enter synchronous meiosis and total RNA was extracted at the indicated times. Samples were processed and blotted, and filters were hybridized with a slk1 probe. As a loading control, 18S rRNA stained with Methylene Blue is shown. exp, exponential growth; meiS, pre-meiotic S-phase; MI, meiosis I; MII, meiosis II. (B) The same experiment as in A, but in this case strain h–/h– pat1-114/pat1-114 slk1-GFP/slk1-GFP (S1706) harbouring a functional version of slk1 tagged with GFP was used, and protein was extracted at the indicated times and processed for western blot. Slk1-GFP was detected with anti-GFP antibodies. Tubulin levels are shown as a loading control. (C) FACS analysis of samples taken during the experiment shown in A. Pre-meiotic S-phase occurs between 1.5 and 2 hours. Spores generated are released during sample processing, and appear as a 1C peak between 6 and 9 hours. (D) Meiosis progression of the experiment shown in A was monitored by DAPI staining. White square, one nucleus; triangle, two nuclei (meiosis I); circle, three to four nuclei (meiosis II); black square, spores. At least 300 cells were counted for each time-point.
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