Figure 5.
52sce-binding proteins co-immunoprecipitate with APP mRNA from a human neuroblastoma cell line. (A) The indicated proteins were immunoprecipitated from SH-SY5Y cytosol fraction (“Input”) with a specific antibody (“IP”) or with pre-immune mouse IgG (“(−)”). (B) mRNA from immunoprecipitates was reverse-transcribed with random primer and subjected to qPCR with APP-specific primers. The amount of APP signal, n = 3, was quantified and normalized to that seen in the pre-immune mouse IgG immunoprecipitate, ± S.E.M. (C) cDNA obtained from immunoprecipitates in a representative experiment in (B) was amplified with primers specific for the APP alternative splice site, and the PCR products were electrophoresed on a 1.5% agarose gel (inverted color scheme). Marker: pGEM DNA size markers (Promega); Input: SH-SY5Y cytosol; RT(−): RT without template; PCR(−): PCR without template. (D) Same set of immunoprecipitates as in (C) was amplified with primers specific for S26, and the PCR products were electrophoresed on a 1.5% agarose gel (inverted color scheme). All lanes are the same as in (C).