Extended Data Fig. 8: Tonoplast ATG8ylation contributes to vacuolar and cellular homeostasis upon cell wall damage.

(a) Vacuolar morphology analyses upon cell wall damage. Representative images and quantification of vacuolar morphology of Col0, ΔCASM and atg16 root cells from the early elongation zone. MDY‐64 (green) staining depicts vacuolar membrane. One-sided Wilcoxon test compared treatments (n = 10) to Col0 wild-type; significant differences (p < 0.01) are indicated as asterisks. The central line indicates the median, and the upper and lower bounds represent quartile 3 (75th percentile) and quartile 1 (25th percentile), respectively. The whiskers denote the minima and maxima of the data points. Scale bar, 10. (b) Transmission Electron Microscopy (EM) images visualizing the vacuole in Col0 and ΔCASM backgrounds under mock and ES20-1 (8 hours, 100 µM) treatments. The images reveal vacuolar fusion between cells in response to cell wall damage. Insets of cell wall damage upon ES20-1 treatment are also included. Representative image from 3 seedlings analyzed under each treatment. Scale bar, 5 µm for the zoom out images and 0,5 µm for the insets of the cell wall. (c) ΔCASM root cells are more sensitive to cell wall damage. Propidium Iodide (PI) staining of root cells from Col0 and ΔCASM backgrounds under mock and ES20-1 (8 hours, 100 µM) treatments, assessing cell viability and membrane integrity. Seven more replicates of Fig. 3f are shown for each genotype and treatment. Scale bar, 10 µm.