Extended Data Fig. 4: Expression patterns of AtICA1, AtICA2, AtICA3, and AtICA4 in Arabidopsis.
a, GUS reporter expression driven by AtICA promoters in Arabidopsis seedlings. Representative images showing GUS staining patterns in whole seedlings of transgenic lines expressing ProAtICA1:GUS, ProAtICA2:GUS, ProAtICA3:GUS, ProAtICA4:GUS, ProAtICA5:GUS, and ProAtICA6:GUS. Scale bars, 1 mm. b, GUS expression patterns in various tissues of mature plants. GUS staining was performed on 20-day-old plants and 40-day-old flowers grown in soil, with observations shown for the entire plant (first column), leaf (second column), and flowers (third and fourth columns). Scale bars are 1 cm (first column), 0.5 cm (second column), 1 cm (third column), and 200 µm (fourth column). c,d, Subcellular localization of AtICA-YFP fusion proteins in roots. YFP fluorescence signals were captured in the root tips (c) and mature root zone (d) of 7-day-old transgenic seedlings expressing ProAtICA1:AtICA1-YFP, ProAtICA2:AtICA2-YFP, ProAtICA3:AtICA3-YFP, and ProAtICA4:AtICA4-YFP. Scale bar, 100 µm (c), 50 µm (d). Experiments were repeated three times with similar results. e, Experimental setup for flg22 treatment for qRT-PCR analysis of AtICA1 expression. Schematic illustrating the treatment of 7-day-old WT seedlings grown in liquid 1/2 MS medium, followed by transfer to liquid 1/2 MS medium supplemented with 200 nM flg22 for the indicated time points. f, AtICA1 transcript abundance is induced by flg22. qRT-PCR analysis of AtICA1 transcript levels in WT seedlings at various time points following treatment with 200 nM flg22, as described in (e). g, Experimental setup for NaCl treatment for qRT-PCR analysis of AtICA1 expression. Schematic illustrating the treatment of WT seedlings grown on solid 1/2 MS medium for 7 days, followed by transfer to solid 1/2 MS medium containing 85 mM NaCl for the indicated time points. h, AtICA1 transcript abundance is induced by NaCl. qRT-PCR analysis of AtICA1 transcript levels in WT seedlings at various time points following transfer to medium containing 85 mM NaCl, as described in (g). Data are mean ± s.d. n = 3. P values were determined using two-tailed Student’s t-tests and indicate significant differences between 0 hour and other time pionts.
