Fig. 2: OsTSN1 interacts with SRBSDV P6 to form droplets.

a A silver-stained SDS-PAGE gel showing P6 co-immunoprecipitated products. Black boxes indicate unique protein bands in SRBSDV-infected rice plants. b Results of RT-qPCR assays showing the transcription levels of OsTSN1 and OsTSN2 in roots, stems, and leaves of rice. Data are the means ± SD from five independent biological replicates, determined using a two-tailed Student’s t test. ****P < 0.0001. c A BiFC assay result showing the interaction between the P6 and OsTSN1 but not OsTSN2 in N. benthamiana leaf cells. Scale bar, 20 μm. d Y2H assay showing the interaction between P6 and OsTSN1 or OsTSN2. Yeast cells co-transformed with pGBKT7-T and pGADT7-P53 were used as a positive control, while cells co-transformed with pGBKT7-T and pGADT7-Lam were used as a negative control. e Pull-down assay showing the interaction between the P6 and OsTSN1. In this assay, GST-P6 was used to pull down MBP-OsTSN1. The membranes were probed with an anti-GST or an anti-MBP antibody. f Predicted structures of the C-P6b (551–793 aa) domain and the OsTSN1-SN3 domain using AlphaFold2 (AF2, https://alphafold.ebi.ac.uk/). The two predicted interaction sites are shown. g Y2H assay results showing the interaction site between P6 and OsTSN1. Yeast cells co-transformed with pGBKT7-T and pGADT7-P53 were used as a positive control, while the cells co-transformed with pGBKT7-T and pGADT7-Lam were used as a negative control. h Subcellular localization of OsTSN1 in rice protoplasts isolated from wild-type (WT) rice or OE-P6 transgenic plants. White arrows indicate droplets. Scale bar, 5 μm. i In vitro co-localization of purified GFP-OsTSN1 and RFP-P6 recombinant proteins. Purified GFP and RFP recombinant proteins served as negative controls. Scale bar, 10 μm. Experiments in (a, c, e, h, and i) were repeated three times with similar results.