Fig. 1.

Developmental regulation of Nuc1 location during meiosis. (A) Nuc1 localization assessed at 0, 6, and 10 h postinduction of sporulation in fixed cells expressing Nuc1-GFP and mito-RFP (mt-RFP). Cells were stained with DAPI to visualize nuclei and mitochondrial nucleoids. (Scale bar, 4 μm.) (B) Anti–Nuc1-FLAG Western blots of mitochondrial (M) and concentrated cytosolic (C) cell fractions sampled at the indicated time points from a synchronized sporulation timecourse of a strain expressing Nuc1-FLAG. A schematic depiction of meiotic progression at each time point is shown above the corresponding Western blots. Fractions were probed with antibodies against Por1 and mAb 414 to validate successful enrichment of mitochondrial and cytosolic contents, respectively. Molecular weight markers are given on the far right. (C) Quantification of Nuc1 abundance in cytosolic and mitochondrial cell fractions at 0 and 6 h postinduction . Data points represent mean of biological replicates (n = 4 per time point) ± SD. Statistical significance was determined by 2-tailed Student’s t test (**P < 0.01). (D) Anti–Nuc1-FLAG Western blots of cell fractions sampled at meiotic time points from wild-type and a mutant ndt80Δ/Δ strain expressing Nuc1-FLAG (fractionation controls included). Strains used: MEY322, MEY783, MMY7415, MMY7599, and SCY53.