FIG. 5.

NELF complex binds to RNA. (A) The indicated amounts (in nanograms) of GST-E(C) WT or 6 μl of Flag-NELF containing 30 ng of Flag-E were incubated with either the HIV TAR or the G-free RNA probe (∼0.1 pmol) with or without one of two different anti-NELF-E monoclonal antibodies (1 μg) for 30 min at 30°C. Reactions were analyzed by electrophoresis on 4% native polyacrylamide gels at 4°C. In lanes 8 to 13, the film was exposed for a longer time. (B and C) The same protein samples containing indicated amounts (in nanograms) of GST-E(C) WT or Flag-E were analyzed by SDS-PAGE and either Coomassie blue staining (B) or immunoblotting (C) with the anti-NELF-E antibodies used in panel A.