Fig. 4: SRBSDV P6 co-localizes with SGs via its interaction with OsTSN1. | Nature Communications

Fig. 4: SRBSDV P6 co-localizes with SGs via its interaction with OsTSN1.

From: Viral proteins suppress rice defenses by boosting OsTSN1 RNA decay via phase separation and multimerization

Fig. 4: SRBSDV P6 co-localizes with SGs via its interaction with OsTSN1.

a Subcellular co-localization of P6, PBs, and SGs in the agroinfiltrated N. benthamiana leaf cells at 2 days post agroinfiltration (dpa). The expressed NbDCP1-CFP and NbG3BP-mCherry were used as a PB marker and an SG marker, respectively. Scale bar, 20 μm. b Percentage of P6 or P6-OsTSN1 droplets colocalized with PBs and SGs in (a). Error bars represent SD, values are means ± SD (n = 12) from three independent biological experiments. c Subcellular co-localization of P6 and SGs in TRV-GUS- or TRV-NbTSN- inoculated N. benthamiana leaf cells, or in the TRV-NbTSN- inoculated N. benthamiana leaf cells expressing flag-NbTSN1 or flag-OsTSN1. NbG3BP-mCherry was used as an SG marker. In this experiment, we co-expressed GFP-P6 and NbG3BP-mCherry, GFP-P6, NbG3BP-mCherry and flag-NbTSN1, and GFP-P6, NbG3BP-mCherry and flag-OsTSN1 in leaves of N. benthamiana plants at 8 d post TRV-GUS or TRV-NbTSN inoculation. Two days post co-inoculation, the N. benthamiana plants were heated at 42 °C for 30 min followed by confocal microscopy observation. The resulting data were analyzed using the Coloc2 of ImageJ software (1.53 v). Rr represents the Pearson Coefficiency. Scale bar, 20 μm. d The percentage of colocalized and non-colocalized granules of P6 and SGs in different assayed leaves in (c). Values are means ± SD (n = 12) from three independent biological experiments, determined using the one-way ANOVA with Tukey’s test. ns, no significant statistical difference, ****P < 0.0001. e A schematic diagram showing the procedure of SG isolation from N. benthamiana leaves and the isolated SGs. Fluorescence signal from the isolated SGs marked by NbG3BP-mCherry was captured under a confocal microscope. Scale bar, 10 μm. The schematic diagram was created in BioRender. Zeng, M. (2025) https://BioRender.com/dqw0hn8. f Western blot results showing the accumulation of P6 in isolated SGs from various samples described in (c). Experiments in (a, c, and f) were repeated three times with similar results.

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