FIG. 6.

nrc-1 is required for phosphorylation of MAK-2 during conidial germination. (A) DIC image of germinating conidia from wild-type (RLM 40-27), Δmak-2, and nrc-1 strains 8 h postinoculation and from the nrc-1 strain 24 h postinoculation. Arrows indicate fusion events observed in RLM 40-27. Hyphal fusion was not evident in nrc-1, although hyphae frequently made contact or grew over each other (24 h). Bar, 30 μm. (B) Conidia from nrc-1 were inoculated on a cellophane membrane layered onto plates containing Vogel's minimal medium with dextrose at 24°C (nrc-1 is an invertase mutant). Total protein (30 μg) from nrc-1 mycelia was harvested at the 0-, 4-, 8-, 12-, 16-, 20-, and 24-h time points. Protein extracts were separated by SDS-10% polyacrylamide gel electrophoresis and blotted onto nitrocellulose membranes. (Top panel) Blot probed with anti-phospho p44/42 antibodies (PhosphoPlus antibody kit; Cell Signaling Technology). (Bottom panel) Blot from the panel above stripped and probed with anti-p44/42 antibodies. The experiment was repeated three times with identical results.