Fig. 4: RxLR23KM specifically interacts with NbERD15La.

A Y2H confirmed that RxLR23KM interacted with NbERD15La in yeast. Yeast transformants were first grown on SD/-Trp/-Leu (DDO), and then selected on SD/-Trp/-Leu/-His/-Ade/X-α-gal (QDO/X) for activating X-α-galactosidase activity. The images were photographed at 4 days after incubation. B Co-IP was used to examine the interaction of RxLR23KM with NbERD15La in N. benthamiana leaves. Left panels confirm transient expression (+) RxLR23KM-GFP and NbERD15La-FLAG. Equal protein loading is indicated by Ponceau staining (Ponceau S). Right panels show that RxLR23KM-GFP is specifically combined with NbERD15La-FLAG. C RxLR23KM uniquely interacts with NbERD15La upon Co-IP assay. Left panels confirmed transient expression (+) RxLR23KM-GFP and NbERD15La/b/c-FLAG. Equal protein loading was indicated by Ponceau staining (Ponceau S). Right panels showed that RxLR23KM-GFP was specifically combined with NbERD15La-FLAG. D NbERD15La specifically interacts with RxLR23KM but not with its two allelic variants by Y2H. The images were photographed at 4 days after incubation. E The interaction of NbERD15La-FLAG with RxLR23KM-GFP and its two allelic variants was detected using Co-IP in N. benthamiana leaves, respectively. Left panels confirmed transient expression of (+) RxLR23KM-GFP, two allelic variants, and NbERD15La-FLAG alone. Equal protein loading was indicated by Ponceau staining (Ponceau S). Right panels showed that NbERD15La-FLAG was specifically combined with RxLR23KM-GFP. F BiFC showed RxLR23KM specifically interacted with NbERD15La in the nucleus and cytoplasm. Different pairs of constructs were co-expressed in N. benthamiana. The fluorescence was observed and imaged by confocal microscopy at 48 hpi. Scale bars, 20 μm. Each experiment was repeated at least three times. Source data are provided as a Source Data file.