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Fig. 2. Cis- and trans-acting mutations cause derepression of an ade6⁺ reporter gene inserted next to mat3-M. (A) Fivefold serial dilutions of each strain were spotted onto non-selective media (left panel) and on selective media lacking adenine (right panel). Strain PJ78 with a truncated allele at the endogenous ade6 locus, denoted ade6-DN/N, and strain PJ282 with a wild-type allele, ade6⁺, were used as reference strains. The strains shown are: PJ644 (w.t. ade6-M210), PJ654 (w.t. ade6-DN/N), PJ579 (IR-L ${Delta}$ /IR-R ${Delta}$ ), PJ577 (h^K repressed) and PJ586 (h^K derepressed), PJ653 (clr4 ${Delta}$ ), PJ668 (swi6 ${Delta}$ ) and PJ651 (dcr1 ${Delta}$ ). (B) RT-PCR analysis of the transcriptional levels from the ade6⁺ reporter gene at mat3-M performed on total RNA from the same strains as in Fig. 3A, except for the control strains PJ78, PJ282. The lower bands in the top panel are the transcripts from the endogenous ade6-DN/N, and served as an internal control for all strains. The top bands are transcripts from the ade6⁺ reporter gene inserted at mat-3M amplified using the same primers. The numbers below the top panel denote the level of ade6⁺ gene expression in the mating-type region calculated as a ratio between the ade6⁺ reporter gene PCR-product and the ade6-DN/N PCR-product. The lower panel shows control reactions in the absence of reverse transcription (labelled –).

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