Description
Identify mRNA targets of the RNA-binding protein HuR in vivo during Schwann cell development using RIP-chip analysis. IP protocol of endogenous mRNA-transfected HuR complexes was performed as described in Keene et al. (2006). In brief, 500 mg of whole-cell lysate obtained from a pool of NB or P5 sciatic nerves from C57BL6J mice were incubated with a suspension of Protein Sepharose beads (Sigma-Aldrich), pre-coated with 15 mg of either IgG1 (BD Pharmingen) or anti-HuR (Santa Cruz Biotechnology) antibodies. mRNAs were isolated using the phenol-chloroform method.