Boxed regions are magnified in Fig 1B. expressing Pab2-YFP (green), Red1-mCherry (red) and CFP-Mmi1 (blue) were examined. Boxed regions are magnified in Fig 1D. Scale bars: 5 m.(PDF) pgen.1008598.s001.pdf (527K) GUID:?40942B1A-36D2-4BA8-AAE9-A2AAC2C0A0D8 S2 Fig: Red1(196C245) is defective in Rrp6 foci formation. (A) Expression levels of truncated Red1 proteins. Cell extracts were prepared from exponentially growing cells expressing wild-type or truncated Red1-YFP in liquid YE medium and immunoblotted with anti-GFP antibody. -tubulin was used as a loading control. The asterisks indicate non-specific bands. (B) Localization of Rrp6 and Red1 in cells. cells expressing Rrp6-YFP (green) and Red1-mCherry (magenta) from the respective endogenous loci were observed. Dotted lines indicate the shape of cells. Boxed region is usually magnified in Fig 2C. Scale bar: 5 m. (C) Localization of Rrp6, Red1 and Mmi1 in cells. cells expressing Rrp6-YFP (green), Red1-mCherry (red) and CFP-Mmi1 (blue) were examined. Boxed region is usually magnified in Fig 2D. Scale bar: 5 m.(PDF) pgen.1008598.s002.pdf (1.8M) GUID:?131A75B9-E96B-4788-B388-BFD93BBBB48D S3 Fig: Red1(196C245) is defective in meiotic transcript degradation. (A) Growth Ubiquitin Isopeptidase Inhibitor I, G5 profiles of wild-type (and cells. Ten-fold serial dilutions of cells were spotted on YE medium and incubated at the indicated temperatures. (B) Expression of mRNA and mRNA in wild-type (and cells. Transcripts were quantified by RT-qPCR and normalized to 0.05; *** 0.001 compared with the wild-type strain (Students mRNA, mRNA, mRNA, PROMPT and PROMPT in wild-type, and cells. Transcripts were quantified by RT-qPCR and normalized to 0.05; ** 0.01; *** 0.001 compared with the wild-type strain Ubiquitin Isopeptidase Inhibitor I, G5 (Students PROMPT and PROMPT in wild-type (cells. Transcripts were quantified by RT-qPCR and normalized to 0.05; ** 0.01; *** 0.001 compared with the wild-type strain (Students cells expressing Red1, Rrp6-GFP, Mmi1, or chimeric proteins composed of Rrp6, GFP, and full-length or truncated Mmi1 from plasmids. Ten-fold serial dilutions of cells were spotted on MM medium and incubated at the indicated temperatures. (B) Expression of mRNA, mRNA, and PROMPT in cells expressing Red1, Rrp6-GFP, Mmi1, or chimeric proteins composed of Rrp6, GFP, and full-length or truncated Mmi1 from plasmids. Transcripts were quantified by RT-qPCR and normalized to 0.01; *** 0.001 compared with cells carrying empty vector (Students cells expressing Red1, Rrp6-GFP, Mmi1, or chimeric proteins composed of Rrp6, GFP, and full-length or truncated Mmi1 from plasmids. Ten-fold serial dilutions of cells were spotted on MM medium and incubated at the indicated temperatures. (D) Expression of mRNA and mRNA in cells expressing Red1, Rrp6-GFP, Mmi1, or chimeric proteins composed of Ubiquitin Isopeptidase Inhibitor I, G5 Rrp6, GFP, and full-length or truncated Mmi1 from plasmids. Cells were produced in liquid MM medium at 25C and shifted to 37C for 4 hours. Transcripts were quantified by RT-qPCR and normalized to 0.01; *** 0.001 compared with cells carrying empty vector at 37?C (Students mRNA, mRNA, and PROMPT in cells expressing Red1, Rrp6-GFP, Mmi1 or chimeric proteins composed of Rrp6, GFP, and full-length or truncated Mmi1 from plasmids in liquid MM medium. Transcripts of each gene were analyzed by RT-qPCR and normalized to 0.05; ** 0.01 compared with cells carrying empty vector (Students cells Ubiquitin Isopeptidase Inhibitor I, G5 expressing Red1, Rrp6-GFP, Mmi1, or chimeric proteins composed of Rrp6, GFP, and full-length or truncated Mmi1 from plasmids. Ten-fold serial dilutions of cells were spotted on MM medium and incubated Rabbit Polyclonal to OR1A1 at the indicated temperatures. (C) Expression levels of chimeric Rrp6-YFP-Mmi1 proteins. Cell extracts were prepared from exponentially growing cells expressing Rrp6-YFP or chimeric proteins composed of Rrp6, YFP, and full-length or truncated Mmi1 from plasmids in liquid MM medium and immunoblotted with anti-GFP antibody. -tubulin was used as a loading control. (D) Expression of mRNA, mRNA and.