Fig 2.

Heterokaryotic ΔR4 strain. (A) ΔR4 strain grown in the selective medium MMC (pH 4.5) in triplicate and incubated at 30°C for 5 days. The circles indicate a white colony and a yellow colony. The numbers of spores per cm² from white and yellow colonies were determined. Plugs of agar (1 cm²) were removed in triplicate for spore calculation. (B) PCR performed with genomic DNA from the wild-type strain (wt), a yellow colony (yc), and a white colony (wc) with the primers pkaR4F-Pr (lanes 1, 4, and 7) or pkaR4F-pkaR4R (lanes 2, 5, and 8). Tef-1f-Tef-1r was used as an internal control (lanes 3, 6 and 9). The primers are shown in Fig. 1E. (C) RT-PCR performed with RNA from wt, dc and wc with primers pkaR4F-pkaR4R, with Tef-1f/Tef-1r as internal control. The positions and sizes of the DNA fragments are indicated at the left of the panels. The quantification of the PCR products, after normalization with the tef-1 signal, is shown in the right panel. The dark bars represent the mutant allele, and the white bars represent the wt allele. The primers used are shown in Fig. 1E.