FIG. 8.

Effects of SUMO1 conjugation of S-HDAg on HDV G-RNA, AG-RNA, and mRNA synthesis. HDV AG-RNA (A) or HDV G-RNA (B and C) was cotransfected with an mRNA encoding wild-type S-HDAg (wt), SUMO1-S-HDAg (SUMO-HDAg), or defective S-HDAg (d1nt) into cells to establish HDV RNA replication. At day 4 (A and B) or day 1 (C) posttransfection, total RNAs were extracted, and HDV G-RNA (A), AG-RNA (B), and mRNA (C) were detected by qRT-PCR, using the G-S, AG-S, and mI-S protocols, respectively (48) (upper panels). The data were normalized relative to the values obtained for the wild-type S-HDAg-encoding mRNA cotransfection. Error bars represent standard deviations from the means for three independent experiments. To demonstrate the expression of the expected forms of HDAg (as indicated by arrows), the whole-cell extracts from one of the qRT-PCR assays were prepared and subjected to immunoblotting with a mouse monoclonal antibody to HDAg or SUMO1, as indicated (lower panels).