Additional file 6: of Insulin-like growth factor receptor and sphingosine kinase are prognostic and therapeutic targets in breast cancer

OSI-906 inhibits p-IGF1R and IGF1R signaling factors in breast cancer cell-lines. 3 × 105 ER-positive A. MCF7 and ER-negative B. HCC-1806 breast cancer cells were plated per well of a 6-well plate and cultured for 24 h and subsequently treated with the dual IGF1R/InsR dual tyrosine kinase inhibitor (OSI-906; 0.1, 0.4 and 1.6 μM) and/or SphK1 inhibitor (SKI-II; 4 μM) for 24 h. Protein lysates were collected and 20 μg of protein was used for immunoblot analysis to measure changes to IGF1R signaling (i.e. AKT and eIF4E-BP1 (protein translation)) and SphK1 steady-state protein expression levels. B-actin was used as a loading control. Abbreviations: Hyper-P = hyper-phosphorylation and Hypo-P = hypophosphorylated. (DOCX 255 kb)