Figure 6.

The Accumulation Level of 66K Affects Viral RNA Replication.

Arabidopsis protoplasts were infected with TYMV RNA and were either treated with DMSO or MG132 (lanes 1 and 2) or cotransfected with water (lanes 3 and 6) or with 1 or 10 μg of the expression vector encoding 66K (lanes 7 and 8) or control protein (REL, Renilla luciferase; lanes 4 and 5) as indicated. Cells were collected at 48 hpi. Equivalent protein amounts were subjected to immunoblot analysis using anti-66K antibody, and equivalent RNA amounts were subjected to strand-specific RNA gel blot analysis to detect (−)-strand and (+)-strand gRNA and sgRNA TYMV RNAs. Methylene blue staining of the membrane (rRNA) is shown to indicate RNA loading.