Extended Data Fig. 5: Requirement of IGFBP motif in modulating IGF-1R/IRS signaling.
From: Insulin-like growth factor-binding protein-7 (IGFBP7) links senescence to heart failure
(a) Diagrams representing 6xHis-tagged WT (ad-IGFBP7) and mutant IGFBP7 (ad-ΔSP, ad-ΔIGFBP, ad-ΔKazal and ad-ΔIgLD) adenovirus (Ad) constructs. (b,c) AC16 cells were infected with Ad-hIGFBP-6xhis for 48 hours. (b) Representative immunoblot shows IGF-1R and IR binding adaptor protein IRS1 and IRS2 were pulldown by IP with anti-6xHis Dynabeads. (c) Representative immunoblot shows IGFBP7 were pull down by reverse IP with anti-IR antibody; The co-IP poll-down bands were indicated by red arrows. (d,e) AC16 cells were infected with Ad-hIGFBP7 (WT), its mutant forms and control (ad-GFP) for 48 hours. (d) Representative immunoblot shows deletion of IGFBP motif, reduced IGFBP7 co-IP with IGF-1Rβ. (e) Representative immunoblot shown ad infected 6xHis-tagged hIGFBP7 and its mutant forms were readily detected by immunoblot with anti-6His antibody as indicated by red cycle; and deletion of IGFBP motif, diminished AKT phosphorylation. Red cycle indicated down regulation of phosphor-Akt. GAPDH was used as loading control.
