Association of L13a and its mutant variants with 28S rRNA. Lysates prepared from HEK 293T cells expressing HA-tagged L13a (WT or mutant as indicated above each lane) were used for immunoprecipitation with anti-HA-coated agarose beads (or nonantibody-coated blank beads as a control, first lane from left). (Top) Total RNA was extracted from the immunoprecipitates and analyzed by RT-PCR with primers specific for 28S rRNA. RT-PCR products were visualized on an ethidium bromide-stained agarose gel. (Bottom) Equal volumes of the immunoprecipitate from each reaction were run on an SDS-PAGE gel and immunoblotted with anti-HA antibody to confirm the presence of equivalent amounts of HA-tagged protein.
