Journal: Nature Communications

Article Title: ATG8ylation-mediated tonoplast invagination mitigates vacuole damage

doi: 10.1038/s41467-025-62084-3

Figure Lengend Snippet: a An overview of the chemical structures of monensin sodium, nigericin sodium, and salinomycin sodium. b The formation of membrane-like structures of GFP-ATG8a in response to three ionophores. Scale bar, 20 μm. c The impact of different concentrations of monensin on the subcellular localization of YFP-ATG8e. 5-day-old YFP-ATG8e transgenic seedlings were treated with different concentrations of monensin (0, 5, 10, 20, 40 μM) in 1/2MS liquid medium for 1 h, followed by confocal microscopy observation. Scale bar, 20 μm. d Time series analysis of the dynamics of GFP-ATG8a in response to monensin (20 μM). Time was presented in minutes. Scale bar, 10 μm. e – i Colocalization analyses between ATG8 and the tonoplast marker VAMP711-mCherry ( e ), the late endosome marker Rab7-GFP ( f ), the early endosome marker YFP-ARA7 ( g ), the TGN marker VHA-a1-RFP ( h ), as well as the cis -Golgi marker GFP-SYP32 ( i ). Scale bars in e – i , 20 μm. j Quantification of the colocalization ratios shown in ( e – i ). The Pearson correlation (PSC) coefficient was analyzed by ImageJ with the PSC colocalization plugin. The data represent means ± SD. n = 6 confocal images (112.5 µm × 112.5 µm) of individual roots. Significance analysis using unpaired two-sided Student’s t test. Similar confocal imaging results were obtained from at least six individual roots, with three replicates. Source data are provided as a Source Data file.

Article Snippet: The ionophores, including monensin sodium salt (MCE, #HY-N0150), salinomycin sodium salt (MCE, #HY-17439), and nigericin sodium salt (MCE, #HY-100381), were prepared as 10 mM stock solutions in ethanol and stored at − 20 °C.

Techniques: Membrane, Transgenic Assay, Confocal Microscopy, Marker, Imaging