Figure 2: RNA Immunoprecipitation (IP). SK-N-SH cells lysates were incubated with protein A/G agarose beads labeled with antibodies to either hnRNP A1 (ab4791) or IgG (Millipore 12-370). Proteins and any bound RNA partners were eluted from the beads. (A) Protein eluents were run on 10% Tris-glycine gels for Western blotting, probed for hnRNP A1 protein (Millipore 4B10 05-1521). L=Untouched SK-N-SH lysate, A1=hnRNP A1 bound agarose beads eluent, IgG=IgG bound agarose beads eluent. The blots show that hnRNP A1 was present in the lysate (L) and the anti-hnRNP A1 IP (A1) but not in the control IgG IP (IgG). (B) RNA was isolated from protein eluent for each group and amplified by RTPCR (see methods). Values within groups were normalized to GAPDH. Results revealed that hnRNP A1 and SPG4 are RNA binding partners of hnRNP A1 and SPG-7 showed a positive trend towards RNA binding. *p ≤ 0.05 analyzed by t-test