Fig. 2.
A cartoon that outlines the RiboTag methodology. The RiboTag mouse is crossed to any available Cre recombinase-expressing mouse line, which results in the deletion of wild-type exon 4 and replacement with the HA-tagged exon 4 only in cells that express Cre recombinase. The cells within the tissue or organ that now express RPL22ha-tagged ribosomes are homogenized, and HA antibody-coupled magnetic beads are added to the cleared homogenate. After an overnight incubation at 4 °C, the magnetic beads that have immunoadsorbed polysomes are washed with a high salt buffer before extraction of the mRNA transcripts, which can then be analyzed by qRT-PCR, microarray, or RNA-seq.