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. 2019 May 23;10:2292. doi: 10.1038/s41467-019-10274-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Suppression of AvrPm3-Pm3 recognition by SvrPm3^a1/f1. a-c Suppression of the recognition of AvrPm3^a2/f2 (a), AvrPm3^b2/c2 (b) and AvrPm3^d3 (c) in presence of the active allele of the SvrPm3^a1/f1 suppressor (right spot). Ratios of Avr-R-Svr mixes for AvrPm3^a2/f2, AvrPm3^b2/c2 and AvrPm3^d3 were 3:1:4, 4:1:4, and 2:1:4 respectively. HR is compared to a negative control in presence of the inactive suppressor allele svrPm3^a1/f1 (left spot). Results are consistent over at least two independent assays each consisting of at least 10 independent leaf replicates. HR quantification in presence of the inactive vs. active suppressor is depicted in the lower panels. Number of independent leaf replicates is indicated. Complete N. benthamiana leaf pictures are provided in Source Data File. d Western blot detection of C and N terminal HA and FLAG epitope fusions of the active SVRPM3^A1/F1 and Ponceau staining of the membrane (lower panel) are depicted. e Western blot detection HA-SVRPM3^A1/F1 and HA-svrPM3^A1/F1 suppressors demonstrating there is no difference in protein abundance between active vs. inactive alleles, respectively. Uncropped Western blot images are provided in Source Data File. f,g Functionality of tagged HA-svrPm3^a1/f1 and HA-SvrPm3^a1/f1 (g) compared to non-tagged svrPm3^a1/f1 and SvrPm3^a1/f1 (f). HR is induced by combining AvrPm3^a2/f2 and Pm3f ^L456P/Y458H. Suppression is assessed by comparing active vs. inactive suppressors, as originally described in Bourras et al.²⁸. Avr-R-Svr ratios are indicated. HR was measured using HSR imaging 3 days post N. benthamiana agro-infiltration (f-g, upper panel). HR Quantification from the same assays revealed by HSR imaging (f-g lower panels). In both cases, the presence of the active suppressor allele (SvrPm3^a1/f1) resulted in significant reduction of HR as compared to the inactive alleles (svrPm3^a1/f1) independently of the epitope, demonstrating that HA-svrPm3^a1/f1 and HA-SvrPm3^a1/f1 are functional. Number of independent leaf replicates is indicated. Statistical significance in a-c, f, g was assessed with a two-sided Student t-Test for paired data and indicated with (*; p < 0.05). Mean values are indicated by the middle line in the boxplot. Individual data points are plotted along the whiskers delineating minimum and maximum values. Raw data underlying the reported averages in a-c, f, g are provided in a Source Data File