Fig. 3: | Nature Communications

Fig. 3

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

Fig. 3

Ime1 and Ume6 control IRT2 expression. a Schematic overview of the IME1 locus indicating the position of the Ume6-binding site (left). Nucleotide coordinates of the Ume6 site relative to IME1 AUG in different Saccharomyces species (right). b Binding of Ume6 to the IME1 promoter measured by chromatin immunoprecipitation using V5 tagged Ume6 (FW2978). A wild-type (FW1509) and an Ume6-binding deletion mutant (pIME1-Δu6, FW2000) strains were also included in the analysis. The means ± SEM of n = 3 experiments are shown. c Northern blot of IRT2 expression in cells that were pre-grown in rich medium and pre-sporulation medium, before shifted to SPO. Wild-type (FW1511), single, and double mutants harboring pIME1-Δu6 and/or a 3′ end mutation in IME1 (ime1-t) (FW2449, FW2370, and FW2571) strains were used for the analysis. To control for loading, the membrane was also probed for SCR1. d IRT1, IRT2, and IME1 expression detected by northern blot in cells expressing IME1 from the copper-inducible promoter (pCUP-IME1, FW3006). pIME1-Δu6 (FW2842) cells were also included in the analysis. Cells were pre-grown in rich and pre-sporulation medium before shifted to SPO and were either not treated (−Cu) or treated with copper sulfate (+Cu) at 1 h in SPO. Northern blot membranes were hybridized with a probe that detects both IRT1 and IRT2, and a probe that detects IME1. As a loading control, the ribosomal RNA is displayed