Fig. 1: Ionophores induce the translocation of ATG8 to the tonoplast.

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 ttest. Similar confocal imaging results were obtained from at least six individual roots, with three replicates. Source data are provided as a Source Data file.