Figure 3.

Immunocapture of TBSV dsRNA and associated proteins. (A) Northern blot analysis of high molecular weight RNA to detect TBSV genomic RNA. RNA was extracted from total (left) and anti-GFP immunoprecipitated (right) fractions of 35S:GFP and 35S:B2:GFP/Col-0 A. thaliana systemically infected with TBSV. A non-infected Col-0 control was included among the total fractions, to the far left. (B) Northwestern blot analysis of the RNA samples described in (A), size-separated on a non-denaturing agarose gel, to detect double-stranded RNA through recombinant B2-Strep. In (A) and (B) ethidium bromide staining of the gels was used as loading control. (C) Northern blot analysis of low molecular weight RNA from the samples described in (A), to detect TBSV-derived siRNA. As loading control, the same membrane was separately hybridized to probes specific to miR159 and snU6. (D) Western blot analysis of proteins from the samples described in (A) to detect GFP. Here, the non-infected Col-0 present in (A–C) was not included. (E) Volcano plot representation shows the enrichment of proteins from TBSV-infected plants that co-purified with B2:GFP. Y- and X-axis display adjusted p-values and fold changes, respectively. The dashed line indicates the threshold above which proteins are significantly enriched (adjP  <  0.05). The source data are available in Supplementary Table S3.