Extended Data Fig. 3: Pir1 degradation by Ubi4 and Cul4–Ddb1 impacts gene silencing.
a, Western blot analysis of GFP–Pir1 in tor2+ cells carrying pir1-WT, pir1-SD or pir1-SA allele. Cells were cultured in rich medium at 30 °C; SD, serine285 replaced by aspartic acid; SA, serine285 replaced by alanine. b, Western blot analysis of Pir1 in strains grown at 30 °C with or without nitrogen (+N/–N). c, IF of Pir1 in the indicated strains at 30 °C. d, Serial dilutions of the indicated strains were spotted onto rich medium (YEA) and low adenine (YE) plates, and then incubated at 30 °C for 3 days. The spd1∆ was introduced to suppress the growth defect of cul4∆ and ddb1∆ mutants. e, RNA-seq expression profiles of sme2 and mei2 transcripts in the indicated strains at 30 °C. f, Serial dilutions of the indicated strains were spotted and incubated as described in d. g, RNA-seq analysis of MTREC regulon gene expression in the indicated homothallic strains at 30 °C. For a-g, data representative of two independent experiments are shown. Source data are provided.
