Figure 3.
UV-cross-linking to determine the molecular weight of RNA binding proteins to domain V of the CVB3 IRES. (α32P)-CTP-labeled probes were added to BHK-21 cell total protein extracts and cross-linked by UV-light exposure, followed by RNase A treatment. Proteins that cross-linked to radioactive RNAs were detected by 10% SDS-PAGE and subsequent autoradiography. Lanes DV CVB3 (Wt) and DV CVB3 (S3) demonstrate protein-probe interactions within CVB3 wild-type and Sabin3-like domain V RNAs, respectively. Lane DV FMDV indicates RNA-protein complexes formed within the FMDV domain V. Equal amounts of total proteins (10 μg) were loaded per well. The molecular weight of each complex was determined by comparing to a concurrently loaded molecular weight (MW) marker: Prestained SDS-page standards broad range (Biorad).