rie1 mutants accumulate lower levels of Ime1 protein. (A–C) Strains harboring N-terminal sfGFP-tagged (sfGFP-IME1) and rie1∆ (B1653, red), wild type RIE1 (B1662, blue), rie1-F240L (B2403, yellow), rie1-F298L (B2836, pink), or rie1-F604L (B2298, purple) were induced to sporulate at 30°C. (A) Protein levels of Ime1 and Pgk1 (loading) were determined by immunoblot (IB), and mRNA levels of IME1 and rRNA (loading) were determined by Northern blot. (B) Quantifications of Ime1 protein corrected for mRNA levels are shown. Biological replicates = 5. (C) Examples of images quantified for Fig. 2 E. sfGFP-Ime1 is shown in green and DAPI is shown in blue. Scale bar, 5 µm. Source data are available for this figure: SourceData FS3.