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. 2024 Sep 1;15:7608. doi: 10.1038/s41467-024-51962-x

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 1 — a Interaction between IPI7 and IPA1 in the yeast two-hybrid assay. IPA1 protein was fused with the GAL4 binding domain to generate BD-IPA1, and IPI7 with the GAL4 activation domain to form AD-IPI7. Blue clones in an X-Gal assay and clones grown on the SD-L-T-U medium indicate protein interaction in yeast cells. b GST pull-down assay for interaction between IPI7 and IPA1. MBP-IPI7, but not MBP, was pulled down with GST-IPA1 immobilized on glutathione-agarose beads. The immunoblot was probed separately with MBP and GST antibodies. Similar results were obtained from three independent biological experiments. c BiFC assay for interaction between IPI7 and IPA1 in tobacco leaves. IPA1 was fused with cYFP (C terminus of YFP) and IPI7 with nYFP (N terminus of YFP). Yellow fluorescence indicates interaction between IPI7 and IPA1 in the nucleus. DAPI was used as the nuclear marker. Bars = 10 μm. Similar results are obtained from three independent biological experiments. d IPI7 interacts with IPA1 in vivo. Total proteins from the protoplasts expressing IPI7-HA or GFP-HA were IP’d with a HA antibody. Proteins before (input) and after IP were detected with an antibody against IPA1 or HA. Similar results are obtained from two independent biological experiments.