Fig. 5.

Asexual spores from ΔsakA and ΔatfA mutants show decreasing viability and lack SakA.

A. Conidia from strains CLK43 (WT), TOL1 (ΔsakA) and TFLΔatfA-02 (ΔatfA) were collected from 5-day plates, counted and plated immediately (time 0) or maintained in water at 4°C for up to 21 days. At indicated time points, aliquots were diluted and used to inoculate supplemented MM plates, which were incubated at 37°C for 2 days and resulting colonies were counted. Data are mean values from three independent experiments; bars indicate standard deviation.

B. Freshly collected conidia or mycelial samples from strains indicated in (A) were frozen with liquid nitrogen and used to prepare total protein extracts, followed by immunoblotting (50 µg), with anti-Hog1 and anti-Phospho-p38 or anti-tubulin antibodies as reported (Kawasaki et al., 2002).

C. Total RNA isolated from intact or germinated conidia from strain CLK43 (WT) was used for northern hybridization with probes against atfA or the 18S ribosomal RNA gene as a loading control.