Profiling Kif5A RNA targets in sensory neuron axons

We previously identified that the RNA binding protein Nucleolin is localized to axons of DRG sensory neurons by interaction with the molecular motor Kinesin-1 (Kif5A) and subsequently localizes importin beta1 mRNA there (Perry et al., 2016). To further identify additional RNAs that are localized to axons in a similar mechanism, we immunopercipitated Kif5A from dorsal roots (centrally projecting axons) or Sciatic nerves (peripherally projecting axons), isolated the bound RNA and sequnced it. We conducted Kif5A immunopercipitation from axoplasm using a rabbit polyclonal antibody. We used a rabbit isotype control (IgG) and also sequenced some of the input material. Axoplasm was extracted either from Dorsal roots (DR) or Sciatic nerves (SN) of the same animal, and 5 animals were pulled per biological repeat. We repeated this in 3 independent biological repeats.