Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Differential subnuclear localization of the (+)- and (-)-strands of PSTVd in infected tomato tissues, with U14 as nucleolar marker. Localization of Mock-inoculated cell shows no PSTVd hybridization signals. The (+)-PSTVd is localized in the nucleoplasm and nucleolus (green). U14 is localized in the nucleolus (red). In the nucleolus, the (+)-PSTVd colocalizes with U14 as indicated by the yellowish fluorescence in the merged image. The (-)-PSTVd (green) is localized in the nucleoplasm but not in the nucleolus, as indicated by the lack of overlap with U14 (red) in the merger image. DAPI stains the nucleoplasm but not the nucleolus, providing an additional marker for the location of the nucleolus. Bars = 2 μm.
• Supplemental Figure 2 - Serial confocal images of spatial localization patterns of the (+)-strand of PSTVd that are suggestive of various stages of nucleolus-bound traffic of the RNA from the nucleoplasm. These are the same cells as shown in Figure 3 of the text. At stage 1 (S1), the viroid RNA is localized only in the nucleoplasm, as indicated by nonoverlapping of PSTVd and U3 signals throughout the entire series. At S2, there is concentration of the viroid RNA as a nucleoplasmic ring around the nucleolus. At S3, the viroid RNA ring appears within the nucleolus. At S4, the viroid RNA is within the nucleolus. The serial optical section images provided an important indication that the viroid RNA is within and not in the periphery of the nucleolus. The diagrams at the bottom of the figure summarize these patterns. Bar = 2 μm.
• Supplemental Figure 3 - Fluorescence microscopic images of the spatial localization patterns of the (+)-strand of PSTVd that are suggestive of various stages of nucleolus-bound traffic of the RNA from the nucleoplasm. (A) to (D) At stage 1 (S1), the viroid RNA is localized only in the nucleoplasm. (E) to (H) At S2, there is concentration of the RNA as a nucleoplasmic ring around the nucleolus. (I) to (L) At S3, the RNA ring appears within the nucleolus. (M) to (P) At S4, the RNA is within the nucleolus. Bar = 2 μm.