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. 2022 Mar 21;13:1524. doi: 10.1038/s41467-022-29196-6

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© The Author(s) 2022

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 — a–c AVR1-CO39 (a), AVR-Pia (b), and AVR-PikD (c) were injected (black arrows) at 1 µM for 2 min on the different MBP:HMA fusion proteins captured by anti-MBP antibody immobilized on the chip. Superimposed sensorgrams are shown for wild-type RGA5_HMA (green), RGA5_HMAm1 (black), RGA5_HMAm1m2 (blue), Pikp-1_HMA (red), as well as for MBP alone (gray) that serves as a negative control. The binding curves obtained with the wild-type and an inactive variant of AVR1-CO39 (a) or AVR-Pia (b) are shown in the top and lower insets, respectively. d Comparison of the binding response (bound fraction) at 1 µM of AVR effectors, expressed as the percentage of the theoretical maximum response (%Rmax) normalized for the amount of MBP:HMA immobilized on the chip and corrected for the MBP-tag contribution. Bars and error bars represent the mean and average deviation calculated for the %Rmax values estimated from n = 2 independent experiments carried out on two different days (open green and red circles) wit n = 1 to n = 4 technical replicates per experiment. Two independently purified protein samples were used to test the binding of the AVR-PikD effector to the different HMAs.