Journal: Journal of Cellular and Molecular Medicine

Article Title: Elevation of IGF-2 receptor and the possible underlying implications in end-stage heart failure patients before and after heart transplantation

doi: 10.1111/j.1582-4934.2011.01414.x

Figure Lengend Snippet: Western blot analysis of cardiac IGF-2R. (A) Representative Western blot analysis of cardiac IGF-2R expression. (B) Densitometric quantification of cardiac IGF-2R expression. The IGF-2R protein levels related to the internal standard protein GAPDH were calculated as the relative abundance. Densitometric analyses of the blots showed an apparent increase in IGF-2R in DCM failing hearts (DCM-HF, n = 5) compared with non-failing control hearts (NF, n = 5; ** P < 0.01). Data are presented as mean ± S.D.

Article Snippet: Sections were then incubated with primary antibodies for 1 hr at room temperature and washed in PBS buffer for 10 min., followed by incubation with IgG-peroxidase conjugated secondary antibody (Sigma-Aldrich, St. Louis, MO, USA) for 1 hr at room temperature, washed in PBS buffer for 10 min., and incubated with 0.5 mg/ml diaminobenzidine tetrahydrochloride 2-hydrate plus 0.05% H 2 O 2 for 5 min. Primary antibodies used included goat anti-human IGF-2R antibody (2 μg/ml, Cat. No. AF2447; R&D System Inc., MN, USA) and rabbit anti-human granzyme B polyclonal antibodies (1:150, Cat. No. Z2145; ZETA Corporation, CA, USA).

Techniques: Western Blot, Expressing, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Elevation of IGF-2 receptor and the possible underlying implications in end-stage heart failure patients before and after heart transplantation

doi: 10.1111/j.1582-4934.2011.01414.x

Figure Lengend Snippet: Histopathological analysis of IGF-2R. (A) Representative light microscopic findings in haematoxylin and eosin staining. The normal appearance of myocardial fibres with central nuclei is seen in non-failing control hearts (NF) and interstitial fibrosis replacement is found in DCM failing hearts (DCM-HF). (B) Representative light microscopic IGF-2R immunoreactivity using a monoclonal antibody, which recognizes IGF-2R. Few and weak immunoreactivity of IGF-2R is observed in non-failing control hearts (NF), but more and strong IGF-2R immunoreactivity can be seen in DCM failing hearts (DCM-HF). Arrows indicated the positive staining.

Article Snippet: Sections were then incubated with primary antibodies for 1 hr at room temperature and washed in PBS buffer for 10 min., followed by incubation with IgG-peroxidase conjugated secondary antibody (Sigma-Aldrich, St. Louis, MO, USA) for 1 hr at room temperature, washed in PBS buffer for 10 min., and incubated with 0.5 mg/ml diaminobenzidine tetrahydrochloride 2-hydrate plus 0.05% H 2 O 2 for 5 min. Primary antibodies used included goat anti-human IGF-2R antibody (2 μg/ml, Cat. No. AF2447; R&D System Inc., MN, USA) and rabbit anti-human granzyme B polyclonal antibodies (1:150, Cat. No. Z2145; ZETA Corporation, CA, USA).

Techniques: Staining, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Elevation of IGF-2 receptor and the possible underlying implications in end-stage heart failure patients before and after heart transplantation

doi: 10.1111/j.1582-4934.2011.01414.x

Figure Lengend Snippet: ELISA of serum IGF-2R and serum CD8. (A) The mean level of serum IGF-2R in DCM heart failure patients before heart transplantation (Bef-Htx, n = 11) was significantly higher than that in non-failing control subjects (NF, n = 11). After heart transplantation, serum IGF-2R levels further increased, peaked at the first month, and gradually reduced close to the pre-operative level at the 6th months, but remained to be higher than that in non-failing controls. ** P < 0.01 versus NF. ## P < 0.01 versus Bef-Htx. (B) The mean level of serum CD8 in DCM heart failure patients before heart transplantation (Bef-Htx, n = 11) had no increase when compared with that in non-failing control subjects (NF, n = 11). Mean levels of serum CD8 also had no change at different time point after heart transplantation compared with that before heart transplantation (Bef-Htx). Data are presented as mean ± S.D.

Article Snippet: Sections were then incubated with primary antibodies for 1 hr at room temperature and washed in PBS buffer for 10 min., followed by incubation with IgG-peroxidase conjugated secondary antibody (Sigma-Aldrich, St. Louis, MO, USA) for 1 hr at room temperature, washed in PBS buffer for 10 min., and incubated with 0.5 mg/ml diaminobenzidine tetrahydrochloride 2-hydrate plus 0.05% H 2 O 2 for 5 min. Primary antibodies used included goat anti-human IGF-2R antibody (2 μg/ml, Cat. No. AF2447; R&D System Inc., MN, USA) and rabbit anti-human granzyme B polyclonal antibodies (1:150, Cat. No. Z2145; ZETA Corporation, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Transplantation Assay, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Elevation of IGF-2 receptor and the possible underlying implications in end-stage heart failure patients before and after heart transplantation

doi: 10.1111/j.1582-4934.2011.01414.x

Figure Lengend Snippet: Immunohistochemical detection of IGF-2R and granzyme B-positive T lymphocytes. Representative immunohistochemical staining on the serial sections from biopsies with rejection grade 3 from patients #6 (A) and from biopsies with rejection grade 2 from patient #4 (B) at the 3 rd month after heart transplantation showed coexistence of IGF-2R and granzyme B-positive T lymphocytes, accompanied by mononuclear cell infiltration with myocyte damage in the same sites in haematoxylin and eosin (HE) staining.

Article Snippet: Sections were then incubated with primary antibodies for 1 hr at room temperature and washed in PBS buffer for 10 min., followed by incubation with IgG-peroxidase conjugated secondary antibody (Sigma-Aldrich, St. Louis, MO, USA) for 1 hr at room temperature, washed in PBS buffer for 10 min., and incubated with 0.5 mg/ml diaminobenzidine tetrahydrochloride 2-hydrate plus 0.05% H 2 O 2 for 5 min. Primary antibodies used included goat anti-human IGF-2R antibody (2 μg/ml, Cat. No. AF2447; R&D System Inc., MN, USA) and rabbit anti-human granzyme B polyclonal antibodies (1:150, Cat. No. Z2145; ZETA Corporation, CA, USA).

Techniques: Immunohistochemical staining, Staining, Transplantation Assay

Journal: Molecular cancer research : MCR

Article Title: Intrinsic Resistance to Cixutumumab is Conferred by Distinct Isoforms of the Insulin Receptor

doi: 10.1158/1541-7786.MCR-15-0279

Figure Lengend Snippet: Insulin-like growth factor II (IGF-II) mediates resistance to cixutumumab in tumor cells over-expressing IR-B. Clonogenic assay in A549 cells that overexpress IR-B (A) in the presence or absence of cixutumumab and/or anti-IGF-II mAb. Represented are mean values +/− SEM, expressed as percentage of colony formation relative to control treated cells. Statistical significance was determined by one-way ANOVA followed by Tukey’s posthoc analysis. Statistically significant difference vs control treated cells (a), cixutumumab-treated cells (b), anti-IGF-II treated cells (c). (B, C) Western blot analysis of signal transduction in A549-IR-B cells treated with IGF-II (25 nM), in the presence or absence of cixutumumab and linsitinib. Serum-starved cells were pretreated with cixutumumab (100 nM) or linsitinib (0.01, 0.1 or 1 uM) for 15 min and 2 h, respectively, followed by incubation with rhIGF-II (25 nM) for 10 minutes. Proteins (15–20 ug) extracted from cultured cells were size-fractionated by SDS-PAGE and immunoblotted with anti-phospho-IRβY1150/51/IGF-IRβY1135/36, 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. (D) MCF-7 and A549 cells with or without IR-A or IRB overexpression were treated with increasing concentrations of linsitinib (0.00015-10 uM) for 72 h, and tumor cell viability was quantified by CellTiter-Glo assay. The results are expressed as % of inhibition of tumor cell viability.

Article Snippet: The following antibodies were purchased from commercial sources as indicated: mouse monoclonal antibodies against Akt (#2920) and p42/p44 MAPK (Erk1/2) (#9107), rabbit monoclonal antibodies against phospho-Akt S473 (#4060) and phospho-IGF-IR beta Y1135/1136 / Insulin Receptor beta Y1150/1151 (#3024), rabbit polyclonal antibody against phospho-p42/p44 MAPK T202/Y204 (Erk1/2) (#9101) (Cell Signalling, Beverly, MA, USA); mouse monoclonal antibody against IGF-IR (#MS-641-P) and Insulin Receptor (#MS-632-P) (Thermo Fisher Scientific, Fremont, CA, USA); rabbit polyclonal Insulin Receptor (#sc-711) and (#sc-7953) (Santa Cruz Biotechnology, Santa Cruz, CA, USA); mouse monoclonal antibody against IGF-II (MAB292) and goat F(ab') 2 anti-mouse IgG-phycoerythrin (#F0102B) (R&D Systems, Minneapolis, MN, USA); goat polyclonal anti-mouse IRDye 680 conjugated (#926–32220) and anti-rabbit IRDye 800 conjugated (#926–32211) (LI-COR Biosciences, Lincoln, Nebraska, USA).

Techniques: Expressing, Clonogenic Assay, Control, Western Blot, Transduction, Incubation, Cell Culture, SDS Page, Over Expression, Glo Assay, Inhibition

Journal: eLife

Article Title: A role for CIM6P/IGF2 receptor in memory consolidation and enhancement

doi: 10.7554/eLife.54781

Figure Lengend Snippet:

Article Snippet: Antibody , anti-Human IGF-II R (goat polyclonal) , R and D Systems , Cat# AF2447, RRID: AB_442153 , 5 ng/μL or 50 ng/μL.

Techniques: Sequencing, Recombinant, Reverse Transcription, SYBR Green Assay, Software