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. 2019 Jan 30;20(4):575–588. doi: 10.1111/mpp.12776

Figure 3.

Figure 3

Rx1 activation triggers single‐stranded DNA (ssDNA) and double‐stranded DNA (dsDNA) breaks. (A) Terminal deoxynucleotidyl transferase dUTP nick‐end labelling (TUNEL) assay revealing DNA damage in DEX::CP105 and DEX::CP106 agroinfiltrated Rx1:4xHA leaves following dexamethasone (DEX) application. Fluorescein‐labelled UTP is incorporated at sites of DNA damage. To allow sample comparison, the fluorescence of fluorescein (proxy for DNA damage) was divided by that of 4′,6‐diamidino‐2‐phenylindole (DAPI)‐stained DNA (proxy for DNA amount). Each value consists of three to six samples and each sample was obtained from separate plants. Statistical analysis using Wilcoxon non‐parametric test: ns, no significant difference; *P < 0.05. Comet assay showing DNA damage in plant nuclei with induced DEX::CP105 and DEX::CP106 constructs in a 4‐h time course, using (B) high‐alkaline (AN) buffer showing ssDNA and dsDNA breaks, and (C) using high‐salt (NN) buffer revealing only dsDNA breaks. Nuclei (comets) were counted and visualized using a box‐and‐whisker plot. The number of nuclei per sample can be found in Table S1. Statistical analysis using Wilcoxon non‐parametric test: ns, no significant difference; **P < 0.001; ***P < 0.0001.