Figure 5
From: Identification of a set of KSRP target transcripts upregulated by PI3K-AKT signaling
Insulin stabilizes a set of KSRP-interacting mRNAs. (A) Serum-starved HIRc-B cells were treated for 1 h with either PBS (control) or insulin (10-6 M). Total extracts were immunoprecipitated with either anti-AKT antibody or control IgG (cIgG). Pellets were incubated (20 min at 30°C) with histone 2B (H2B) in kinase buffer in the presence of γ[32P]ATP under gentle shaking. Labeled proteins were separated by SDS-PAGE and detected by autoradiography. (B) In vitro RNA degradation assays using S100 extracts from either control or insulin (10-6 M)-treated HIRc-B cells. Internally 32P-labeled, capped RNA substrates (see Additional file 4 for sequences) were incubated with S100 extracts for the indicated times and their decay analyzed as described in Methods.