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. 2016 Dec 23;7:1945. doi: 10.3389/fpls.2016.01945

FIGURE 3.

FIGURE 3

Overexpression of PER57 increases the permeability of the leaf cuticle. (A) Toluidine blue (TB) exclusion assay. Leaves of 21-day-old C24 and ohy1 plants were stained with 6 μl of 0.05 % TB drops at indicated time points (n = 8). (B) TB-stained areas were quantified in Fiji (n = 8, ±SD). ∗p < 0.05, Student’s t-test. (C) C24 and ohy1 plants were treated with CWDEs. CWDEs, when applied to plant tissue, cause symptoms that can include tissue maceration and cell death. Cell wall fortification was assessed 6 and 24 h post-treatment (hpt) with CWDEs (n = 6). (D) Quantitative RT-PCR analysis was performed to evaluate the levels of MYB96, BDG and LACS2.3 transcripts in 28-day-old ohy1 rosette leaves at 24 h after the plants were sprayed with a mock solution or 100 μM ABA. Transcript levels were plotted relative to the expression level in the C24 line at 0-hpt. The EF1α and UBQ10 reference genes were used as internal controls. Error bars represent the standard deviation of three biological replicates. ∗p < 0.05, Student’s t-test. (E) Quantitative RT-PCR analysis was performed to evaluate the levels of CER1, LTP3, LTP4 and KCS2 transcripts in 28-day-old ohy1 rosette leaves at 24 h after the plants were sprayed with a mock solution or 100 μM ABA. Transcript levels were plotted relative to the levels in the C24 line at 0-hpt. The EF1α and UBQ10 reference genes were used as internal controls. Error bars represent the standard deviation of three biological replicates. ∗p < 0.05, Student’s t-test. All experiments were repeated twice with similar results.