MoEnd3 functions in nuclear degradation and autophagy.

(A) Nuclei were visible during appressorium development using H1:RFP. The merged image shows H1:RFP and DIC. Bars = 10 μm. (B) Electron micrographs of vacuoles in hyphae following 4 h nitrogen starvation condition. Arrows indicate autophagosomes. Bars = 0.5μm. (C) Hyphae from the strains expressed GFP:MoAtg8 were exposed to nitrogen starvation for 4 h in the presence of 4 mM PMSF. Arrows indicate vacuoles. Merged image shows the GFP:MoAtg8 and DIC. Bar = 10μm. (D) The percentages of vacuoles containing GFP:MoAtg8 were recorded by observing at least 100 vacuoles for each sample, and the experiment was repeated three times. Error bars represent SD and asterisk represents significant difference (P < 0.01). (E) GFP:MoAtg8 proteolysis assay. Total proteins were extracted from the GFP:MoAtg8 expressed strains exposed to nitrogen starvation condition for 0, 2 and 5 h. The full-length GFP:MoAtg8 and free GFP were detected using GFP antibodies. Protein contents were analyzed using the actin antibody.