Figure 1: Nature Communications

Fig. 1

From: A regulatory circuit of two lncRNAs and a master regulator directs cell fate in yeast

Fig. 1

Transcription of IRT2 promotes entry into meiosis. a Scheme of the IME1 locus consisting of: IRT1, IRT2/MUT1573, and the IME1 gene. b IRT2 expression in SK1 diploid cells (FW1511) during entry into meiosis. Cells were grown in rich medium till saturation, shifted and grown in pre-sporulation medium for another 16 h, and transferred to sporulation medium (SPO). Samples for northern blot were taken at the indicated time points. A probe directed to the upstream region in the IME1 promoter was used to detect IRT2. To control for loading, membranes were also probed for SCR1. c IRT2 expression in S288C diploid cells (FW631) during entry into meiosis. Cells were grown till saturation in rich medium, and subsequently shifted to SPO. Samples were taken at the indicated time points. IRT2 levels were quantified by reverse transcription and quantitative PCR. The signals were normalized to ACT1 levels. The means ± SEM of n = 2 experiments are shown. d Scheme of IME1 and RME1 alleles used in e and f. e Quantification of cells that completed meiotic divisions (MI + MII) in diploid S288C cells that were either wild type (FW631), expressed IRT2 from the inducible CUP1 promoter (pCUP-IRT2) (FW2668) or harbored a deletion of RME1 (rme1Δ, FW1497). Cells were grown as described in c, at 2 h in SPO, pCUP-IRT2 cells were either not treated (−Cu) or treated with copper sulfate (+Cu). Cells were fixed after 72 h in SPO, stained, and DAPI masses of n = 200 cells were counted. Cells with two or more masses were considered to have completed at least one meiotic division. Means ± SEM of n = 5 experiments are shown. ***p < 0.0005; ****p < 0.0001 (Student’s t test). f Quantification of cells that completed meiotic divisions (MI + MII) in SK1 diploid cells that were either wild type (FW1511), pCUP-IRT2 (FW5254), rme1Δ (FW2340), harbored deletions in the a1α2 repressor sites of the RME1 promoter (RME1-H, FW1196), harbored pCUP-IRT2 and rme1Δ (FW2476), or pCUP-IRT2 and RME1-H (FW2385). Cells grown and treated as described in e. Means ± SEM of at least n = 4 experiments are shown. *p < 0.05; ****p < 0.0001 (Student’s t test)