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. 2009 Dec 1;4(12):e8111. doi: 10.1371/journal.pone.0008111

Figure 4. Fun30 is involved in silencing in the HMR locus.

Figure 4

A. Diagram of the HMR silencing reporter (adapted from [32]). B. Colonies of the strains YLS410 (wildtype silencer, [32]), YLS404 (ΔA silencer, [32]) and YLS586 (ΔB silencer, [32]) turn red because of lack of expression of ADE2. When FUN30 is deleted in these backgrounds, the colonies stay white, indicating derepression of ADE2. C. The role of Fun30 in repressing the ADE2 reporter embedded in the HMR is tested by monitoring growth in the absence of adenine. The repression of the ADE2 reporter gene in the ‘wt’ strain (YLS410) leads to reduced growth compared to the fun30Δ isogenic strain. 10-fold serial dilutions of cells were spotted on agar plates with minimal glucose medium (MinGlu) or minimal glucose medium without adenine (MinGlu-Ade) and incubated for 3 days at 30°C. D. Fun30 does not regulate ADE2 in its endogeneous locus. The amount of ADE2 mRNA expressed from its endogenous locus in control (JS311, [37]) and its fun30-deleted sister strain was determined after reverse transcription of the mRNA and quantitative PCR and normalized to ACT1 expression. Shown is the average of two experiments with standard deviation.