Sciatic nerve crush regulation of ribosome-associated RNA in DRG neurons, alternative polyadenylation analytsis [DRGinjuryRiboTag2018-QuntSeq72h]

Nerve injury alters transcription and translational processes in cell-type specific manner.Here we probe the effect of sciatic nerve injury on alternative polyadenylation of ribosome associated RNA specifically in sensory neurons.Mice were subjected to crush of the sciatic nerve in one side, while the other nerve was spared and used as naive control condition. Adv-Cre//Ribotag mice were subjected to crush injury on in one sciatic nerve. Corresponding DRG (Lumbar L4-L6) were extracted 3 days after injury. Injured and naive ganglia was extracted from each animal and subjected to immunoprecipitation (IP) of Ribotag ribosomal marker (HA-tagged Rpl22) to isolate intact ribosomes from sensory neurons. RNA was extracted from the input (prior to immunoprecipitation) and post-IP fraction. 3 replicates were used, combining DRG tissue from 4 mice per repeat for RNA isolation.