Fig. 4: NIb disrupts the function of Pelota by competing with SCE1.

a, b The relationships between the levels of SCE1-Pelota BiFC and NIb-CFP fluorescence intensity in N. benthamiana leaves. Scale bars correspond to 20 μm. Dashed lines separate different levels of NIb fluorescence activity in cells. This experiment was repeated three times showing similar results. Statistical analysis was performed using linear regression to fit the data points. The regression line represents the best fit, and the R² value indicates the goodness of fit. The p value was calculated using a two-sided test, and no adjustments for multiple comparisons were applied. c, Competitive MBP pull-down assays in vitro. MBP-SCE1 protein combined with GST-Pelota was incubated with a gradient amount of His-NIb. The proteins were immunoprecipitated with Amylose Resin. Input and pull-down proteins were analyzed with anti-GST, His, and MBP antibodies. Quantification of protein levels was performed using ImageJ software. Independent experiments were performed three times with similar results. The samples were derived from the same experiment and that blots were processed in parallel. d Competitive Co-IP assays in vivo. SCE1-YFP and Myc-Pelota were co-expressed with a gradient amount of of NIb-Flag in N. benthamiana leaves. Proteins were immunoprecipitated with anti-GFP beads at 40 hpi, and the immunoprecipitated and input proteins were then analyzed with anti-Myc, anti-Flag, and anti-GFP antibodies. CBB-staining of RbcL was a loading control. Quantification of protein levels was performed using ImageJ software. Independent experiments were performed three times with similar results. The samples were derived from the same experiment and that blots were processed in parallel. e Growth phenotype of Col-0, pelota mutant and NIb-OE transgenic Arabidopsis plants. Bar = 5 cm. f The overlap of significantly upregulated genes (log₂ fold change > 2, p < 0.001) in transcriptome analysis between NIb overexpressing (in red) and pelota (in blue) plants. g The PCA plot displays the distribution of transcriptome data from wild-type (WT, green dots), NIb transgenic (red dots), and pelota (blue dots) samples. Statistical significance between the two datasets was assessed using a two-sided Mann–Whitney U test. h Gene ontology terms relating to NIb transgenic and pelota samples compared to WT show enrichment across biological processes, cellular components, and molecular functions. The dot size reflects the enrichment score, and the color gradient indicates the −log₁₀ (p-value), indicating the significance of the enrichment. i Functional annotation and network of enriched terms, illustrated by pie charts colored to reflect the proportions of differentially expressed genes within the NIb transgenic (red) and pelota (blue) groups. GO enrichment analysis was performed using a hypergeometric test. P values were adjusted for multiple comparisons using the Benjamini-Hochberg method to control the false discovery rate (FDR).