Fig. 5.

Salt-induced responses are alleviated by CaCl₂ or EGCG application. (A,B) Seven-day-old seedlings of Col-0, fer-4 and herk1 the1-4 lines were treated for 15 min with water (mock, ‘m’), 100 mM NaCl (‘N’), 5 mM CaCl₂ (‘C’) or NaCl/CaCl₂ (‘NC’) (A) or with DMSO (‘D’), 100 mM NaCl (‘N’), 50 μM EGCG (‘E’) or NaCl/EGCG (‘NE’) (B). MPK phosphorylation was detected by immunoblotting. Equal loading was checked by using the anti-β-actin antibody. Similar results are shown in Fig. S5A,B. (C,D) Expression of PROPEP3, WRKY40 and RRTF1 determined by qRT-PCR was performed in seven-day-old seedlings of the Col-0, fer-4, herk1 the1-4 lines treated for 1 h with water (mock, ‘m’), 100 mM NaCl (‘N’), 5 mM CaCl₂ (‘C’) or NaCl/CaCl₂ (‘NC’) (C) or with DMSO (‘D’), 100 mM NaCl (‘N’), 50 μM EGCG (‘E’) and NaCl/EGCG (‘NE’) (D). n=3. Error bars indicate standard deviation (s.d.). One-way ANOVA and Tukey's HSD (α=0.05) were used to analyze statistical differences within the same genotype (lowercase letters for Col-0, uppercase letters for fer-4, italic letters for herk1 the1-4). Multiple two-tailed, unpaired t-tests according to Benjamini–Hochberg correction between NaCl treatments and NaCl/CaCl₂ treatments are shown. **P<0.01; ***P<0.001. (E) Four-day-old seedlings of the Col-0, fer-4 and herk1 the1-4 lines were subjected to the halotropism assay with DMSO (mock), 200 mM NaCl, 50 μM EGCG or NaCl/EGCG. Histograms represent the average root angle of three independent experiments, error bars show s.e.m. n=75. Different letters indicate statistical significance by one-way ANOVA and Tukey's HSD (α=0.05) within the same genotype (lowercase letters for Col-0, uppercase letters for fer-4, italic letters for herk1 the1-4). Asterisks indicate differences between salt treatments according to multiple two-tailed, unpaired t-tests coupled with Benjamini–Hochberg correction. ***P<0.001; n.s., not significant.