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: Bioscience, biotechnology, and biochemistry

Article Title: Chemiluminescent enzyme immunoassay for measuring leptin.

doi: 10.1271/bbb.100885

Figure Lengend Snippet: Fig. 1. Schematic Representation of the CLEIA Procedure for Leptin.

Article Snippet: According to the manufacturers’ instructions, the measurable ranges for leptin are 15.6–1000 pg/mL for the Human Leptin Immunoassay kit (Qiagen, Hamburg, Germany) and Leptin ELISA Human kit (R&D Systems, Minneapolis, MN, USA), 1–50 ng/mL for the Human Leptin ELISA kit (B-Bridge International, Cupertino, CA, USA), and 0.5–100 ng/mL for the Human Leptin ELISA kit (Linco Research, St. Charles, MO, USA).

Techniques:

Journal: Bioscience, biotechnology, and biochemistry

Article Title: Chemiluminescent enzyme immunoassay for measuring leptin.

doi: 10.1271/bbb.100885

Figure Lengend Snippet: Fig. 5. Standard Curves for Leptin. Leptin (100 mL of 0–1.0 pg/mL) was used to make a standard curve. The CV values at each point were 1.3–5.7% (n ¼ 6). Error bars indicate SD values.

Article Snippet: According to the manufacturers’ instructions, the measurable ranges for leptin are 15.6–1000 pg/mL for the Human Leptin Immunoassay kit (Qiagen, Hamburg, Germany) and Leptin ELISA Human kit (R&D Systems, Minneapolis, MN, USA), 1–50 ng/mL for the Human Leptin ELISA kit (B-Bridge International, Cupertino, CA, USA), and 0.5–100 ng/mL for the Human Leptin ELISA kit (Linco Research, St. Charles, MO, USA).

Techniques:

Journal: Stem cells (Dayton, Ohio)

Article Title: Nuclear fusion-independent smooth muscle differentiation of human adipose-derived stem cells induced by a smooth muscle environment.

doi: 10.1002/stem.1023

Figure Lengend Snippet: Figure 6. Significant upregulation of human HGF in conditioned medium was confirmed by ELISA. HGF promotes smooth muscle differentia- tion of human adipose-derived stem cell (hASC) in a dose-dependent manner. (A): Increase of HGF from undetected (time 0) to more than 1000 pg/ml at 6 weeks differentiation. (B): At the lower concentrations, there was a dose-dependent induction of alpha smooth muscle actin (ASMA) in hASC exposed to HGF. Higher induction was seen at concentrations between 1 and 5 ng/ml of HGF at 6 weeks. All groups are statistically significant when compared to DMEM (p < .05). (C): Immunofluorescence staining confirmed fluorescence-activated cell sorter data. Double fluo- rescence staining visualizing the nuclei (blue) and ASMA (green). Undifferentiated hASCs before injection show 2%–3% baseline ASMA expres- sion (1). After 6 weeks in SMIM medium, there is an increase in the expression of ASMA (2). Further increase in ASMA expression is seen when cells are exposed to 5 ng/ml HGF for 6 weeks (3) (scale bar ¼ 100 lm). Abbreviations: DMEM, Dulbecco’s modified Eagle’s medium; FBS, fetal bovine serum; HGF, hepatocyte growth factor; SMIM, smooth muscle induction medium.

Article Snippet: Test and control media (DMEM or SMIM media exposed to hASC at time 0) were collected and examined for hepatocyte growth factor (HGF) levels using the Quantikine Human HGF ELISA kit (R&D Systems, Minneapolis, MN, www.rndsystems. com) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Staining, Injection, Expressing

Journal: Cancer gene therapy

Article Title: Mc-hES, a novel plasmid carrying human endostatin gene, inhibits nasopharyngeal carcinoma growth.

doi: 10.1038/cgt.2011.72

Figure Lengend Snippet: Figure 2 Expression profiles and biological function of endostatin in eukaryotic cells. (a) RT–PCR analysis of endostatin transcripts in CNE-2 cells transfected with plasmid. Lane 1, DL2000, bands of 2, 1, 0.75, 0.5 and 0.25 kb; lane 2, mc-hES; lane 3, pcDNA-hES; lane 4, pj-hES; lane 5, p2jc31; lane 6, empty control; lane 7, positive control. (b) Western blot assay. Lane 1, p2jc31; lane 2, mc-hES; lane 3, pcDNA-hES; lane 4, pj-hES. (c) Expression profiles of endostatin in the supernatant of cells transfected with plasmids. Columns represent the mean of three independent experiments, each conducted in triplicate; bars, s.e. Po 0.05, compared with pj-hES and pcDNA-hES treated group. (d) MTT assay. HUVECs were treated with CM from transfected CNE-2 cells and cell proliferation was evaluated after 72 h. Columns, cell proliferation normalized against p2jc31 control (mean±s.d.). Mc-hES versus pcDNA-hES, P ¼ 0.019; mc-hES versus pj-hES, P ¼ 0.005; pcDNA-hES versus p2jc31, P ¼ 0.237. (e) Tube formation assay. HUVECs were treated with CM. After 16 h of incubation images of tube formation were captured and tube formation was scored in one 50 microscopic field. The number of tubes formed was quantitated and the data are presented as the mean±s.d. per field ( 50). Data were normalized to p2jc31-treated control (n ¼ 5). Mc-hES versus pcDNA-hES, P ¼ 0.015; mc-hES versus pj-hES, P ¼ 0.008; pcDNA-hES versus p2jc31, P ¼ 0.009. (f) Migration assay. HUVECs were treated with CM and pipetted into inserts of matrigel-coated transwells. Addition of chemoattractant, HUVECs in CM containing 50 ng ml1 vascular endothelial growth factor, to the lower well of boyden chamber stimulated endothelial migration to the underside of the transwell membrane. After 6 h of incubation, migrated cells were stained by DAPI. The number of cells that migrated was counted by microscopy and the data are presented as the mean±s.d. per field ( 100). Data were normalized to p2jc31-treated control (n ¼ 5). Mc-hES versus pcDNA-hES, P ¼ 0.01; mc-hES versus pj-hES, P ¼ 0.015; pcDNA-hES versus p2jc31, P ¼ 0.019.

Article Snippet: CNE-2 cell supernatants were harvested as conditioned medium (CM) after 72 h of transfection and endostatin expression was evaluated using a human endostatin ELISA kit (R&D systems, Minneapolis, MN) according to the recommendation of the manufacturer.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, Control, Positive Control, Western Blot, MTT Assay, Tube Formation Assay, Incubation, Migration, Membrane, Staining, Microscopy

Journal: Cancer gene therapy

Article Title: Mc-hES, a novel plasmid carrying human endostatin gene, inhibits nasopharyngeal carcinoma growth.

doi: 10.1038/cgt.2011.72

Figure Lengend Snippet: Figure 4 (a) Endostatin expression profile in the tumor tissues. For expression profiles of endostatin, results are given in nanograms per mg of tumor tissue after single administration. Columns, mean of three mice; bars, s.d. mc-hES versus pcDNA-hES. Po0.001 at days 3, 7, 14 and 20. (b) Results from representative experiments are shown for RT–PCR analysis of endostatin transcripts. b-actin was used as loading control. b, blank; M, DL2000; , negative control; þ , positive control; s, saline.

Article Snippet: CNE-2 cell supernatants were harvested as conditioned medium (CM) after 72 h of transfection and endostatin expression was evaluated using a human endostatin ELISA kit (R&D systems, Minneapolis, MN) according to the recommendation of the manufacturer.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Negative Control, Positive Control, Saline