Fig. 6: NaCl induces the translocation of AtCLCf from the Golgi to the PM.

a–c Confocal microscopy images of Nicotiana benthamiana leaf epidermal cells show that N-terminal GFP-tagged AtCLCf translocates to the PM when treated with 100 mM NaCl, as indicated by the expression of GFP-AtCLCf along with Golgi, TGN, and PM markers. Shown are GFP-AtCLCf expression (a) before and after (b) 4 h and (c) 6 h of NaCl treatment. The GFP signal was visualized under 488 nm excitation, 500–525 nm emission; the markers were visualized under 561 nm excitation, 575–600 nm emission. Co-localization of GFP-AtCLCf with markers is shown in the merged images. Scale bars, 20 µm, data were obtained from three independent experiments, and representative images are shown here. d Quantification of AtCLCf-GFP localization with respective markers (n = 7) using ImageJ. Horizontal lines within the boxes in d indicate the median. Minimal and maximal data points are indicated by the lower and the upper whiskers, respectively. e Expression of GFP-tagged AtCLCf in the leaf mesophyll protoplasts of pAtCLCf::GFP-AtCLCf Arabidopsis lines without and with NaCl treatment, n = 10. Scale bars, 10 µm. Means with different letters are significantly different, P < 0.05 (one-way ANOVA followed by Tukey’s test).