Supplemental Tables 1-2 & Figures 1-4 from Intrinsic Resistance to Cixutumumab Is Conferred by Distinct Isoforms of the Insulin Receptor

Supplemental Tables 1-2 & Figures 1-4. Supplemental Table 1. Total IR, IR-A, IR-B, IGF-IR, IGF-I and IGF-II mRNA expression in preclinical models of pediatric solid tumors. Supplemental Table 2. Patient-derived xenograft models of lung adenocarcinoma: clinical information and mutational status. Supplemental Figure 1. Effect of cixutumumab on circulating growth hormone (A), insulin (B) and IGF-I (C) levels in male C57BL/6 mice. Represented are mean values +/- SEM. p-values {less than or equal to} 0.05 indicate statistical significance (*) (Student's t-test). Supplemental Figure 2. IGF-I and IGF-II mRNA expression in patient-derived models of lung adenocarcinoma. Represented are mean values +/- SEM. Supplemental Figure 3. Overlay analysis of cell surface IR and IGF-IR expression by flow cytometry (A) and ligand-induced signal transduction by Western Blot (B) in A549 cell overexpressing empty vector (pLVX) or IR isoforms (IR-A and IR-B). Serum-starved cells (1h) were treated with rhIGF-I, rhIGF-II or recombinant human insulin (10 nM) for 5, 15 or 60 minutes. Proteins (10ug) extracted from cultured cells were size-fractionated by SDS-PAGE and immunoblotted with anti-phospho-IRbY1150/51/IGF-IRbY1135/36, anti-phospho-AktS473 and anti-phospho-p42/p44 MAPKT202/Y204 antibodies. Total level of proteins was demonstrated by immunoblotting with antibodies directed against total Akt and p42/p44 MAPK. Supplemental Figure 4. Anchorage-dependent cell viability assay. MCF-7-Mock, MCF-7-IR-A and MCF-7-IRB cells were treated with increasing concentrations of cixutumumab (0.015-1,000 nM). Maximum inhibition for MCF-7-Mock, MCF-7-IR-A and MCF-7-IRB were 60.4%, 0% and 26.9% respectively, p-values < 0.01(Student's t-test). Represented are mean values +/- SEM.