Journal: Journal of Biological Chemistry

Article Title: Silencing of Insulin-like Growth Factor-binding Protein-2 in Human Glioblastoma Cells Reduces Both Invasiveness and Expression of Progression-associated Gene CD24

doi: 10.1074/jbc.m609567200

Figure Lengend Snippet: FIGURE 2. Modulation of IGF/IGF-IR signaling caused by knockdown of IGFBP-2. The IGFBP-2-knockdown cells (IGFBP-2 KD) and control cells were treated with the indicated concentrations of IGF-I or IGF-II in serum-free con- dition, and extents of phosphorylation of IGF-IR (p-IGF-IR), Akt (p-Akt), and ERK1/2 (p-Erk1/2) were analyzed by immunoblot and a representative result of U251 was shown. The band intensity was measured and the ratio of phos- phorylated protein to the corresponding total protein was calculated.

Article Snippet: The following primary antibodies were used: anti-human IGFBP-2 mouse monoclonal antibody (C-10; Santa Cruz Biotechnology, Santa Cruz, CA), anti- -actinmousemonoclonal antibody (AC-74; Sigma), anti-phosphoERK1/2 rabbit monoclonal antibody (Thr185/Tyr187, clone AW39;Upstate Cell Signaling Solutions, Lake Placid, NY), antiphospho-IGF-IR rabbit polyclonal antibody (Tyr1131), antiIGF-IR rabbit polyclonal antibody, anti-phospho-Akt mouse monoclonal antibody (Ser473), anti-Akt rabbit polyclonal antibody, anti-phospho-ERK1/2 rabbit monoclonal antibody (Thr185/Tyr187, clone AW39; Upstate Cell Signaling Solutions, Lake Placid, NY), anti-ERK1/2 rabbit polyclonal antibody, anticaspase-3, and cleaved capase-3 (Asp175) rabbit polyclonal antibodies, and anti-heat shock protein 70 rabbit polyclonal antibody (Cell Signaling Technology, Danvers, MA).

Techniques: Knockdown, Control, Phospho-proteomics, Western Blot

Journal: Cancer Research

Article Title: Chemoresistant Colorectal Cancer Cells, the Cancer Stem Cell Phenotype, and Increased Sensitivity to Insulin-like Growth Factor-I Receptor Inhibition

doi: 10.1158/0008-5472.can-08-2023

Figure Lengend Snippet: Figure 4. Effect of IGF-IR inhibition on in vivo tumor growth, proliferation, and apoptosis. Mice were s.c. injected with 1 106 HT29 or HT29/5FU-R cells in one experiment or HT29 or HT29/OxR cells in a second experiment and treated with control IgG or AVE1642 twice weekly. The final tumor masses were measured and compared between mice bearing tumors from each cell line. Relative to control- treated mice, HT29/5FU-R–derived (A) and HT29/OxR-derived tumors (B) showed significantly greater growth inhibition than did HT29-derived tumors. TUNEL staining revealed significantly greater apoptosis in response to AVE1642 in tumors derived from HT29/5FU-R–derived (C) and HT29/ OxR-derived tumors (D) than in tumors derived from parental cells. Bars, SE. *, P < 0.05.

Article Snippet: Antibodies used for flow cytometry, immunohistochemical analysis, immunofluorescence, or Western blotting were as follows: rabbit anti-CD133, rabbit anti-phosphorylated IGF-IR (Cell Signaling Technology), mouse anti-vinculin, rabbit anti-h-actin (Sigma-Aldrich), mouse anti-CD44 (Abcam), phycoerythrin (PE)-conjugated anti-CD133, PE-conjugated mouse-IgG1 (Miltenyi Biotec), FITC-conjugated anti-CD44, FITC-conjugated mouse-IgG2b, rabbit antiIGF-IR (Santa Cruz Biotechnology), and mouse anti-Ki67 (Dako).

Techniques: Inhibition, In Vivo, Injection, Control, Derivative Assay, TUNEL Assay, Staining