Figure 1

KSRP associates with a set of unstable mRNAs overrepresented in myrAKT1-αT3-1 cells. (A) In vitro RNA degradation assays using S100 extracts from αT3-1 cells. Internally ³²P-labeled, capped RNA substrates (see Additional file 4 for sequences) were incubated with extracts for the indicated times and their decay analyzed as described in Methods. (B) The interaction between ³²P-labeled RNAs (indicated on the right) and recombinant purified KSRP (30–300 nM) was evaluated by UV-crosslinking. (C) Immunoprecipitation of ribonucleoprotein complexes containing different KSRP target mRNAs. The proteins were immunoprecipitated from αT3-1 cell extracts using the indicated antibodies. RNA was extracted from the immune complexes and analyzed by RT-PCR as described in Methods.