Fig. 1

Nuclear translocation of CgCrzA and its interaction with CgMkk1 under CFW stress. A Localisation of CgCrzA under CFW stress was examined by observing mycelia treated with CFW for 60 and 120 min using confocal fluorescence microscopy. Scale bar = 10 μm. B Five grams of mycelia (untreated and treated with CFW for 60 min) was divided into three parts to extract total protein, nuclear protein, and cytoplasm protein. SDS-PAGE was used to separate proteins, followed by western blotting with an anti-GFP antibody to detect CgCrzA. Bands corresponding to CgCrzA-GFP were observed at 102 kDa. CBB, Coomassie Brilliant Blue. C Grayscale analysis was performed on the western blot bands to analyze their intensities. The intensity of CgCrzA was compared between untreated (-) and CFW-treated (+) mycelia (60 min) for total, nuclear, and cytoplasmic proteins. Error bars represent the standard errors from three independent experiments. Asterisks indicate a significant difference at P < 0.01. D Yeast two-hybrid assays for the interaction between CgMkk1 and CgCrzA. The prey and bait constructs were assayed for growth on SD-Leu-Trp and SD-Leu-Trp-His-Ade plates. “+” denotes positive control, while “-” represents negative control. E Co-immunoprecipitation assays for the interaction between CgMkk1 and CgCrzA. Total proteins were isolated from transformants co-expressing CgMkk1-S and CgCrzA-GFP constructs, and proteins eluted from the anti-GFP beads (elution). Immunoblots were incubated with monoclonal anti-GFP and anti-S antibodies