Fig. 3

UvAtg8 is essential for the autophagy in Ustilaginoidea virens. a, Autophagy was blocked in ∆Uvatg8 mutants. The strains were cultured in the PS (potato sucrose medium) medium for 3 d, following subjected to SD-N (nitrogen starvation condition) medium for 4 h to determine the autophagic affuxes in the WT and ∆Uvatg8 strain. The images were taken by bright field microscopy. b, The degradation of the GFP-UvAtg8 was observed under nitrogen starvation condition via Western blot assay with an anti-GFP. GAPDH was used as an internal reference. c, Confocal microscopy image of the GFP-UvAtg8 strain under nitrogen rich and starvation condition. The indicated strain was cultured in the PS medium for 3 d, and then shifted to an SD-N medium for 12 h. The vacuoles were stained with CAMC (7-amino-4-chloromethylcoumarin) before the observation using confocal laser scanning microscope. Scale bar = 5 μm. d, The number of autophagosomes was counted. The data were subjected to Duncan’s Test and the significant difference was indicated by asterisks (***, p < 0.001)