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. 2020 Sep 18;11:570422. doi: 10.3389/fpls.2020.570422

Figure 2.

Figure 2

Evidence that NPR1 autoregulates its own gene induction. (A, B) SA-induced NPR1 mature mRNA (A) and pre-mRNA (B) accumulation in Col-0, npr1-1, and npr1-2. (C) SA-induced NPR1 protein accumulation in Col-0, npr1-1, and npr1-2. (D) SA-induced NPR1 pre-mRNA accumulation in Col-0, npr1-2, and 35S:NPR1-GFP npr1-2. (E) SA-induced NPR1 protein accumulation in Col-0, npr1-2, and 35S:NPR1-GFP npr1-2. The asterisk indicates a band with unknown nature. (F) SA-induced NPR1 pre-mRNA accumulation in Col-0, npr1-3, and NPR1:Myc-NPR1 npr1-3. In (A, B, D, F), 4-week-old soil-grown plants were treated with soil drenches of 1 mM SA solution. Leaf tissues were collected at the indicated time points. Total RNA was extracted and subjected to qPCR analysis. Expression was normalized against the constitutively expressed UBQ5. Data represent the mean of three independent samples ± SD. Different letters above the bars indicate significant differences (P < 0.05, one-way ANOVA in (A, B, D) and Student’s t-test in F). The statistical comparisons were performed among time points for each genotype. In (C, E), 4-week-old soil-grown plants were treated with soil drenches of 1 mM SA solution. Total protein extracted from leaf tissues collected at the indicated time points was analyzed by reducing SDS-PAGE and immunoblotting with anti-NPR1 antibody. Ponceau S staining of RuBisCo confirmed equal loading. All experiments were repeated three times with similar trend.