Fig. 3: HC-ProTu triggers autophagic AGO1 degradation.
From: HC-Pro inhibits HEN1 methyltransferase activity, leading to autophagic degradation of AGO1
a Heatmaps of autophagy-related genes in various P1/HC-Pro plants were generated from transcriptome profiles. The color scale bar indicates the fold change in target gene expression. b Differential expression of ATG8 family genes in various P1/HC-Pro plants relative to their expression levels. Data are presented as mean values ± standard deviations (n = 3). Significant differences were determined by Student’s t-test; * indicates P values < 0.05. ** indicates P values < 0.01. *** indicates P values < 0.001. c Diagram of the gRNA positions in the ATG8a gene (i) and the gene-editing results (ii) for P1/HC-ProTu/atg8age. d ATG8a transcript levels in P1/HC-ProTu and P1/HC-ProTu/atg8age plants compared with those of Col-0. Data are presented as mean values ± standard errors (n = 3). e Hierarchical clustering dendrogram on the transcriptomes of Col-0, P1/HC-ProTu and P1/HC-ProTu/atg8age plants. f Relative expression of autophagy-related genes in P1/HC-ProTu and P1/HC-ProTu/atg8age plants compared to that in Col-0 plants. Data are presented as mean values ± standard deviations (n = 3). Significant differences based on Student’s t-test; * indicates P values < 0.05. ** indicates P values < 0.01. g Evaluation of the suppression efficiency of miRNA-mediated target gene regulation between P1/HC-ProTu and P1/HC-ProTu/atg8age plants by real-time RT-PCR. Relative expression levels were normalized to the ubiquitin level. Data are presented as mean values ± standard errors (n = 6). Significant differences were determined by Student’s t-test; ** indicates P values < 0.01. h Subcellular localization of ATG8a (i), HC-ProTu (ii), and AGO1 (iii) within leaf cells as revealed by immunogold labeling and transmission electron microscopy (TEM). Immunogold signals were detected within and/or surrounding autophagic structures, many of which were encased by autophagic vacuoles. Red arrowheads highlight the presence of immunogold particles within autophagic vacuoles (Av) or autophagic-like structures (Ap), while blue arrowheads denote signals found in the cytoplasm (n = 3). Cw, cell wall; Cv, central vacuole; Ch, chloroplast. Scale bars, 200 nm. i Image size measurements for speckle-like structures. Data are presented as mean values ± standard deviations (n = 10). j FRET efficiency of co-infiltrated plants in which HC-ProTu-CFP or HC-ProTu-K-CFP interacts with YFP-ATG8a. Data are presented as mean values ± standard deviations (n = 9). Source data are provided as a Source Data file.
