Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Effects of individual control of pH and hypoxia in chondrocyte culture.

doi: 10.1002/jor.20994

Figure Lengend Snippet: Figure 2. The pH-dependence of gene expression of chondrocytes in alginate beads. Relative mRNA levels for AGC1, COL1, COL2, SOX9, HIF1A, and VEGF after 5 days of culture were determined by quantitative RT-PCR and normalized to 18S rRNA. Asterisk indicates p < 0.05 (n ¼ 6).

Article Snippet: One hundred microliter of medium was used for VEGF ELISA according to the manufacturer’s instructions (DVE00; R&D Systems, Abingdon, UK).

Techniques: Gene Expression, Quantitative RT-PCR

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Effects of individual control of pH and hypoxia in chondrocyte culture.

doi: 10.1002/jor.20994

Figure Lengend Snippet: Figure 3. Relative VEGF release into the culture medium after 5 days at different pH levels and oxygen tensions, measured by ELISA. Asterisk indicates p < 0.01 (n ¼ 6). Values are normalized to the average of pH 7.4/pO2 5%.

Article Snippet: One hundred microliter of medium was used for VEGF ELISA according to the manufacturer’s instructions (DVE00; R&D Systems, Abingdon, UK).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Biochimica et biophysica acta

Article Title: Fibronectin conformation regulates the proangiogenic capability of tumor-associated adipogenic stromal cells.

doi: 10.1016/j.bbagen.2013.03.033

Figure Lengend Snippet: Fig. 2. Decellularized tumor-associated matrices influence adipogenic stromal cell adhesion and proangiogenic factor secretion. 3T3-L1 cells assemble a denser, more fibrillar matrix with increased Fn content when cultured in the presence of TCM [11], which was reflected with decellularized matrices prepared from control (Control) and TCM-treated 3T3-L1s (Tumor) using detergent-based methods. Scale bar = 20 μm (A). Tumor matrices decreased the total number of adherent 3T3-L1 per well as determined by counting trypsinized cells (B) and increased VEGF secretion per cell as analyzed via ELISA followed by normalization to cell number (C). Varied VEGF levels were not due to differential ECM sequestra- tion because ELISA of lysates of decellularized ECMs did not reveal statistically significant differences between conditions (D). * indicates p b 0.05.

Article Snippet: Subsequently, media was removed and analyzed using a human VEGF ELISA (R&D Systems).

Techniques: Cell Culture, Control, Enzyme-linked Immunosorbent Assay

Journal: Biochimica et biophysica acta

Article Title: Fibronectin conformation regulates the proangiogenic capability of tumor-associated adipogenic stromal cells.

doi: 10.1016/j.bbagen.2013.03.033

Figure Lengend Snippet: Fig. 3. Fn influences cell behavior in response to control and tumor ECMs. Addition of the control peptide Del29 during preparation of tumor ECMs does not alter Fn incorporation, whereas addition of the Fn polymerization inhibitor pUR4 markedly decreased Fn incorporation. Scale bar = 20 μm (A). Accordingly, tumor matrices prepared in the presence of pUR4 increased cell adhesion as determined by counting of trypsinized cells (B) and diminished VEGF secretion as analyzed via ELISA followed by normalization to cell number (C) to levels comparable to control matrices. * indicates p b 0.05.

Article Snippet: Subsequently, media was removed and analyzed using a human VEGF ELISA (R&D Systems).

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Biochimica et biophysica acta

Article Title: Fibronectin conformation regulates the proangiogenic capability of tumor-associated adipogenic stromal cells.

doi: 10.1016/j.bbagen.2013.03.033

Figure Lengend Snippet: Fig. 4. Tumor ECM-induced VEGF secretion by stromal cells stimulates endothelial cell mi- gration. Media collected from 3T3-L1 cells cultured on tumor ECMs increased HUVEC mi- gration in transwell migration assays relative to media collected from control ECMs, as measured via image analysis of DAPI-stained, fixed membranes following migration. Typ- ical micrographs of migrated HUVECs stained with DAPI are shown to the right. Scale bar = 20 μm (A). Blockade of VEGF signaling using a VEGFR2/KDR peptide antagonist confirmed that greater VEGF levels secreted by stromal cells cultured on tumor ECMs con- tributed to changes in HUVEC migration (B). * indicates p b 0.05.

Article Snippet: Subsequently, media was removed and analyzed using a human VEGF ELISA (R&D Systems).

Techniques: Cell Culture, Migration, Control, Staining

Journal: Biochimica et biophysica acta

Article Title: Fibronectin conformation regulates the proangiogenic capability of tumor-associated adipogenic stromal cells.

doi: 10.1016/j.bbagen.2013.03.033

Figure Lengend Snippet: Fig. 5. Conducting polymer films to define the effect of Fn conformation on adhesion and VEGF secretion. Oxidized (+1 V) or reduced (−1 V) PEDOT:PSS surfaces were coated with media containing either fibronectin (Fn) only, or 10% serum (containing Fn), to es- tablish compact and partially unfolded conformations of adsorbed Fn, respectively. The number of 3T3-L1 cells per well adhered to unfolded Fn was decreased relative to compact Fn (A), while an opposite effect was detected for VEGF secretion (B) for both Fn and serum-coated devices. These differences were determined by image analysis of DAPI-stained fixed devices and VEGF ELISA followed by normalization to cell number. * in- dicates p b 0.05.

Article Snippet: Subsequently, media was removed and analyzed using a human VEGF ELISA (R&D Systems).

Techniques: Polymer, Staining, Enzyme-linked Immunosorbent Assay

Journal: Biochimica et biophysica acta

Article Title: Fibronectin conformation regulates the proangiogenic capability of tumor-associated adipogenic stromal cells.

doi: 10.1016/j.bbagen.2013.03.033

Figure Lengend Snippet: Fig. 6. Increased VEGF levels in response to partial Fn unfolding are not due to differential binding to Fn and are relevant to additional cell types. Incubation of cell-free, serum- coated devices with recombinant VEGF followed by ELISA of non-bound VEGF in the media suggested that VEGF preferentially interacts with partially unfolded Fn and, hence, confirms that varied secretions rather than differential binding were responsible for the differences in VEGF levels detected above (A). Similar to 3T3-L1, human bone marrow-derived MSCs enhanced secretion of VEGF when seeded onto substrates with partially unfolded Fn relative to compact Fn as examined via VEGF ELISA followed by nor- malization to cell number (B). * indicates p b 0.05.

Article Snippet: Subsequently, media was removed and analyzed using a human VEGF ELISA (R&D Systems).

Techniques: Binding Assay, Incubation, Recombinant, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: Biochimica et biophysica acta

Article Title: Fibronectin conformation regulates the proangiogenic capability of tumor-associated adipogenic stromal cells.

doi: 10.1016/j.bbagen.2013.03.033

Figure Lengend Snippet: Fig. 7. Varied Fn conformation regulates adhesion and VEGF secretion via altered integrin engagement. Individual and combined blockade of synergy-dependent (α5β1) and -independent (αvβ3) integrins via β1- and αv-function blocking antibodies, respectively, suggested that α5β1 inhibits VEGF secretion of 3T3-L1 in response to compact Fn, which may be compensated by increased engagement of αvβ3 (white bars). Additionally, αvβ3 stimulates VEGF secretion in response to partially unfolded Fn, which is independent of α5β1 usage (black bars). * indicates p b 0.05.

Article Snippet: Subsequently, media was removed and analyzed using a human VEGF ELISA (R&D Systems).

Techniques: Blocking Assay

Journal: PLoS ONE

Article Title: Combined Inhibition of Epidermal Growth Factor Receptor and Cyclooxygenase-2 Leads to Greater Anti-tumor Activity of Docetaxel in Advanced Prostate Cancer

doi: 10.1371/journal.pone.0076169

Figure Lengend Snippet: Cells grown in 1% FBS were exposed to 0.01 μmol/L docetaxel (D), 20 μmol/L gefitinib (G) or100 μmol/LNS-398 (N), alone or in combination, or with DMSO as control for 24 h. NF-KB, MMP-9 and VEGF mRNA (A and B) and protein levels (C and D) were measured as described in Materials and Methods. Values are reported as the mean±SD of three independent experiments.

Article Snippet: Antibodies against human VEGF and MMP-9 were obtained from R&D System and Bioworld, respectively.

Techniques: Control

Journal: Molecular Oncology

Article Title: Focal adhesion kinase inhibitors are potent anti‐angiogenic agents

doi: 10.1016/j.molonc.2011.10.004

Figure Lengend Snippet: FAK inhibitors decrease number of viable endothelial cells in a dose‐dependent manner. HUVEC were incubated with various concentrations of either PF‐228 (A) or FI14 (B) in the presence of VEGF. Cells treated with vehicle (DMSO) were used ...

Article Snippet: Recombinant human vascular endothelial growth factor (VEGF) (rhVEGF 165 ; R&D Systems, Minneapolis, MN) was reconstituted according to the manufacturer's instructions.

Techniques: Incubation

Journal: Molecular Oncology

Article Title: Focal adhesion kinase inhibitors are potent anti‐angiogenic agents

doi: 10.1016/j.molonc.2011.10.004

Figure Lengend Snippet: FAK inhibitors induce cell cycle arrest and apoptosis in treated endothelial cells. Seeded HUVEC were treated with DMSO as vehicle control or varying concentrations of PF‐228 or FI14 in the presence of 50 ng/mL VEGF for 48 h. ...

Article Snippet: Recombinant human vascular endothelial growth factor (VEGF) (rhVEGF 165 ; R&D Systems, Minneapolis, MN) was reconstituted according to the manufacturer's instructions.

Techniques: Control

Journal: Molecular Oncology

Article Title: Focal adhesion kinase inhibitors are potent anti‐angiogenic agents

doi: 10.1016/j.molonc.2011.10.004

Figure Lengend Snippet: FAK inhibitors block HUVEC sprouting on collagen I gels. For assessing formation of sprouts, HUVEC were seeded onto collagen I gel‐coated plates and treated with 50 ng/ml VEGF in the absence (with added DMSO as a vehicle control) ...

Article Snippet: Recombinant human vascular endothelial growth factor (VEGF) (rhVEGF 165 ; R&D Systems, Minneapolis, MN) was reconstituted according to the manufacturer's instructions.

Techniques: Blocking Assay, Control