Journal: Clinical and Vaccine Immunology : CVI

Article Title: Interactive Changes between Macrophages and Adipocytes ▿ †

doi: 10.1128/CVI.00494-09

Figure Lengend Snippet: Influence of peritoneal macrophages on the levels of adiponectin ( ACDC ), GLUT1 , and GLUT4 transcripts in 3T3-L1 adipocytes a

Article Snippet: A rabbit polyclonal antibody against mouse GLUT1 (catalog no. CBL242) was obtained from BD Biosciences (San Jose, CA).

Techniques:

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Interactive Changes between Macrophages and Adipocytes ▿ †

doi: 10.1128/CVI.00494-09

Figure Lengend Snippet: Influence of cytokines on the regulation of the ACDC, GLUT1, and GLUT4 genes in 3T3-L1 adipocytes a

Article Snippet: A rabbit polyclonal antibody against mouse GLUT1 (catalog no. CBL242) was obtained from BD Biosciences (San Jose, CA).

Techniques: Cell Culture

Journal: Clinical and Vaccine Immunology : CVI

Article Title: Interactive Changes between Macrophages and Adipocytes ▿ †

doi: 10.1128/CVI.00494-09

Figure Lengend Snippet: Effects of recombinant TNF-α, IL-6, and IL-1β on GLUT4 protein expression. Fully differentiated adipocytes were incubated in medium containing recombinant mouse TNF-α (2 ng/ml), IL-6 (10 ng/ml), or IL-1β (20 ng/ml) for 2 days. Cells were fixed and immunostained for GLUT1 or GLUT4 expression using flow cytometry as described in Materials and Methods. The data are presented as means ± standard errors of the means for 3 to 6 independent replicates per treatment. Different letters indicate a significant difference. A P-value of <0.05 was considered significant.

Article Snippet: A rabbit polyclonal antibody against mouse GLUT1 (catalog no. CBL242) was obtained from BD Biosciences (San Jose, CA).

Techniques: Recombinant, Expressing, Incubation, Flow Cytometry

Journal: Endocrinology

Article Title: Localization of glucokinase-like immunoreactivity in the rat lower brain stem: for possible location of brain glucose-sensing mechanisms.

doi: 10.1210/endo.141.1.7234

Figure Lengend Snippet: FIG. 3. Glucose transporter-like immunoreactivities in GK-like immunopositive ependymocytes and endothelial cells. The GLUT2-like im- munoreactivities were detected on the cilia of GK-like immunopositive ependymocytes composing the wall of the central canal (A), the aqueduct (B), the third ventricle (C), the lateral ventricle (D), and the fourth ventricle (E). A three-dimensional image of whole mount preparation constructed from the confocal laser scanning microscopic sections revealed that GLUT2-like immunoreactivities (white) were located along the wall of the fourth ventricle, but not on the dorsal surface of area postrema (F). Precise localization of the GLUT2 (black) on the cilia was revealed by the immunoperoxidase histochemistry (G). GLUT2-like immunoreactivity was eliminated by preincubation of anti-GLUT2 antibody with

Article Snippet: We used a rabbit polyclonal antibody to GLUT1 (1:100; Santa Cruz Biotechnology, Inc.) and GLUT4 (1:25; Chemicon International, Inc., Temecula, CA).

Techniques: Construct

Journal: Endocrinology

Article Title: Localization of glucokinase-like immunoreactivity in the rat lower brain stem: for possible location of brain glucose-sensing mechanisms.

doi: 10.1210/endo.141.1.7234

Figure Lengend Snippet: FIG. 6. The schematic representation of the distribution of GK-like immuno- reactive cells in the brain. In addition to the schema, GK-like immunopositive ependymocytes showed the GLUT2-like and GLUT1-like immunoreactivities on the cilia and the GLUT4-like immuno- reactivity predominantly in cytoplas- mic area (see Fig. 3, A–E). Endothelial cells also showed the GLUT1-like im- munoreactivity lining the wall of mi- crovessel (see Fig. 3J). 3V, Third ven- tricle; 4V, fourth ventricle; AP, area postrema; AQ, aqueduct; CC, central canal; DRn, dorsal raphe nucleus; Hip, hippocampus; LV, lateral ventricle; MRn, median raphe nucleus; RMg, ra- phe magnus; RO, raphe obscurus; RP, raphe pallidus; RPo, raphe pontis; Sep, septum.

Article Snippet: We used a rabbit polyclonal antibody to GLUT1 (1:100; Santa Cruz Biotechnology, Inc.) and GLUT4 (1:25; Chemicon International, Inc., Temecula, CA).

Techniques:

Journal: EMBO Molecular Medicine

Article Title: Endothelial cell‐derived angiopoietin‐2 is a therapeutic target in treatment‐naive and bevacizumab‐resistant glioblastoma

doi: 10.15252/emmm.201505505

Figure Lengend Snippet: A, B Immunofluorescence staining of CD 31 and desmin in GL 261 glioblastoma after single and dual treatment with anti‐Ang‐2 ( AMG 386) and anti‐ VEGF (aflibercept) (A). Corresponding quantitative analysis of microvessel densities and pericyte numbers (B; control n = 24; AMG 386 n = 10; aflibercept n = 12; AMG 386 + aflibercept n = 13). C, D Staining for mouse IgG (red) as a surrogate parameter for vascular leakage and CD 31 (green) is shown in GL 261 glioblastoma sections following different anti‐angiogenic treatments (C) with quantification in (D); n = 3. E Quantification of necrotic areas in GL 261 glioblastoma sections after anti‐angiogenic treatment is shown as percent of whole tumor area (control, aflibercept; AMG 386 + aflibercept n = 4; AMG 386 n = 3). F Analysis of Glut1 immunoreactivity in GL 261 glioblastoma sections ( n = 3). G, H Double‐immunofluorescence stainings of macrophages (F4/80) and tumor vessels ( vWF ) in mouse GBM after treatment with AMG 386, aflibercept, or the combination of both are shown in (G). Quantitative analysis of tumor‐infiltrating leukocytes (F4/80 + , CD 3 + , Ly‐6G + ) following anti‐angiogenic treatment is displayed in (H). (Control n = 28; AMG 386 n = 12; aflibercept n = 11; AMG 386 + aflibercept n = 12). I Kaplan–Meier survival analysis of GL 261 tumors grown in C57 BL /6 mice following anti‐angiogenic treatment (control n = 38; AMG 386 n = 13; aflibercept n = 12; AMG 386 + aflibercept n = 13). J, K Double‐immunofluorescence stainings with anti‐F4/80 and anti‐ CD 206 in brain tumor sections of mice treated with anti‐Ang‐2 ( AMG 386), anti‐ VEGF (aflibercept), or the combination of both are shown in (J). Corresponding quantitative analyses of tumor‐infiltrating F4/80 + cells and CD 206 + cells, and the ratio of CD 206 + versus F4/80 + upon anti‐angiogenic therapy is displayed in (K) (control n = 21; AMG 386 n = 13; aflibercept n = 9; AMG 386 + aflibercept n = 4). L, M Flow cytometry of tumor‐infiltrating macrophages following enzymatic dissociation of mouse GL 261 brain tumors plus/minus anti‐angiogenic therapy. Percent of CD 206 + (L) and MHC class II HI cells (M) among CD 45 + CD 11b + GR 1 − F4/80 + cells is displayed (control n = 4; AMG 386 n = 4; aflibercept n = 5; AMG 386 + aflibercept n = 5). Data information: Statistical analyses were performed using one‐way ANOVA and Tukey's multiple comparison except for log‐rank and Wilcoxon (I). * P < 0.05, ** P < 0.01, *** P < 0.005, # P < 0.005 of aflibercept + AMG 386 versus aflibercept; data are mean ± SEM (A–K), mean ± SD (L, M). Scale bars (A, C, G and J): 100 μm.

Article Snippet: Anti‐mouse CD31 (clone MEC 13.1) 1:100, anti‐mouse Ly‐6G (clone 1A8) 1:100, anti‐mouse CD3e (clone 145‐2C11) 1:100 all from BD Pharmingen; anti‐mouse F4/80 (clone A3‐1) 1:100 from Biozol; goat anti‐mouse CD206 (clone C068C2) 1:100 from R&D Systems; anti‐αSMA (clone 1A4) 1:200 from Sigma; anti‐mouse Glut1 (polyclonal, C‐Terminus) 1:100 from Merck Millipore; anti‐human Ang‐1 (Santa Cruz, sc‐6319); and anti‐mouse Ang‐2 (R&D Systems, AF7186).

Techniques: Immunofluorescence, Staining, Control, Flow Cytometry, Comparison