Journal: Animals : an open access journal from MDPI

Article Title: piggyBac Transposition and the Expression of Human Cystatin C in Transgenic Chickens.

doi: 10.3390/ani11061554

Figure Lengend Snippet: Figure 4. Quantification and antimicrobial activity test of hCST3 in transgenic chickens. (A) Quantification of hCST3 in the egg white (EW) of transgenic hens by ELISA. hCST3 was detected in all of the transgenic hen eggs. (B) hCST3 was detected in the muscle of hCST3 transgenic chickens by Western blotting. (C) His-tagged hCST3 in muscle and egg white of hCST3 transgenic chickens was purified and detected by Western blotting after purification with Ni-NTA magnetic nanobeads. (D) Biofunctional activity of hCST3 from transgenic hen eggs. The purified hCST3 from transgenic hen egg white was transferred to Whatman paper disks at different concentrations: (1) control (ddH2O), (2) elution buffer, and (3) 10, (4) 25, (5) 50, (6) 75, and (7) 100 ng hCST3/disc. Original western blot figures in Figure S1.

Article Snippet: hCST3 protein in cells and transgenic chickens was detected and the concentration was determined by using human cystatin C Quantikine® ELISA Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Activity Assay, Transgenic Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: PLoS ONE

Article Title: CD4-Independent Human Immunodeficiency Virus Infection Involves Participation of Endocytosis and Cathepsin B

doi: 10.1371/journal.pone.0019352

Figure Lengend Snippet: (A) A blasticidin resistance gene-bearing mNDK HIV-1 vector was inoculated into the cDNA expression library-transfected HeLa cells, and blasticidin-resistant cell clones were isolated. Transduction titers of the LacZ gene-bearing mNDK HIV-1 vector were measured on these blasticidin-resistant cell clones. Relative values to the transduction titer on normal HeLa cells are indicated. This experiment was repeated 3 times, and the results are shown as the means +− SD. Asterisks indicate statistically significant differences compared to the relative value on HeLa cells. (B) PCR was performed using genomic DNA samples prepared from the HmN-2 cells. The PCR products were subjected to agarose gel electrophoresis. (C) A cell lysate prepared from the HmN-2 cells was analyzed by Western immunoblotting using an anti-cystatin C (upper panel) or anti-actin (lower panel) antibody. (D) To detect cystatin C or GAPDH mRNA, RT-PCR was performed. The PCR products were subjected to agarose gel electrophoresis.

Article Snippet: To construct cystatin C-expressing cells, HeLa, TE671, 293T, and their CD4-expressing cells were inoculated with an MLV vector encoding cystatin C (Origene, Rockville, MD) as previously reported .

Techniques: Plasmid Preparation, Expressing, Transfection, Clone Assay, Isolation, Transduction, Agarose Gel Electrophoresis, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: CD4-Independent Human Immunodeficiency Virus Infection Involves Participation of Endocytosis and Cathepsin B

doi: 10.1371/journal.pone.0019352

Figure Lengend Snippet: (A) Transduction titers of the mNDK HIV-1 vector were measured on cystatin C-transduced and -untransduced HeLa, TE671, 293T, and their CD4-expressing cells. Relative values to the transduction titer on cystatin C-untransduced cells are indicated (upper panel). This experiment was repeated 3 times, and the results are shown as the means +− SD. Asterisks indicate statistically significant differences compared to the relative value on the each cystatin C-untransduced cells. Cystatin C expression was confirmed by Western immunoblotting using the cystatin C antibody (lower panel). (B) The cell surface expression of CXCR4 on the cystatin C-transduced and –untransduced HeLa cells was analyzed by a fluorescence flow cytometer. Closed and open areas indicate cells treated with control serum and then with the secondary FITC-conjugated anti-rat IgG antibody and cells treated with the CXCR4 antibody and then with the secondary antibody, respectively. Relative values to means of fluorescence intensity in the untransduced cells are also indicated. This experiment was repeated 3 times, and the results are shown as averages +− SD.

Article Snippet: To construct cystatin C-expressing cells, HeLa, TE671, 293T, and their CD4-expressing cells were inoculated with an MLV vector encoding cystatin C (Origene, Rockville, MD) as previously reported .

Techniques: Transduction, Plasmid Preparation, Expressing, Western Blot, Fluorescence, Flow Cytometry

Journal: PLoS ONE

Article Title: CD4-Independent Human Immunodeficiency Virus Infection Involves Participation of Endocytosis and Cathepsin B

doi: 10.1371/journal.pone.0019352

Figure Lengend Snippet: (A) Cathepsin B activities in living 293T, TE671, and HeLa cells were analyzed by the cathepsin B detection reagent. The closed area and green line indicate cells unstained and cells stained with the cathepsin B detection reagent, respectively. (B) Fluorescence intensities of cell lysates prepared from HeLa, TE671, 293T, and their CA-074Me-treated cells were measured to quantitate cathepsin B activity by using a cathepsin B activity assay kit. This experiment was repeated 3 times, and the results are shown as the means +− SD. Asterisks indicate statistically significant differences compared to the fluorescence intensity on DMSO-treated control cells. (C) Cathepsin B activities of cell lysates prepared from HeLa, TE671, 293T, and their cystatin C-expressing cells were analyzed by using a cathepsin B activity assay kit. Relative values to the fluorescence intensity for each cell type not expressing cystatin C are indicated. This experiment was repeated 3 times, and the results are shown as the means +− SD. Asterisks indicate statistically significant differences compared to the relative value on the cells not expressing cystatin C.

Article Snippet: To construct cystatin C-expressing cells, HeLa, TE671, 293T, and their CD4-expressing cells were inoculated with an MLV vector encoding cystatin C (Origene, Rockville, MD) as previously reported .

Techniques: Staining, Fluorescence, Activity Assay, Expressing

Journal: Netherlands Heart Journal

Article Title: The ethnicity-specific association of biomarkers with the angiographic severity of coronary artery disease

doi: 10.1007/s12471-015-0798-y

Figure Lengend Snippet: Baseline characteristics of UNICORN participants, stratified by ethnicity

Article Snippet: Commercial antibody combinations were used to quantify NTproBNP (15 C4 and biotinylated 13G12, Hi-test Finland), hsCRP (Dy1707 duoset, R&D systems), CysC (Dy1196 duoset, R&D systems) and MPO (Dy3667 duoset, R&D systems).

Techniques: Medications, Biomarker Discovery

Journal: Netherlands Heart Journal

Article Title: The ethnicity-specific association of biomarkers with the angiographic severity of coronary artery disease

doi: 10.1007/s12471-015-0798-y

Figure Lengend Snippet: Biomarker levels stratified by indication for angiography and by CAD severity, for each ethnicity

Article Snippet: Commercial antibody combinations were used to quantify NTproBNP (15 C4 and biotinylated 13G12, Hi-test Finland), hsCRP (Dy1707 duoset, R&D systems), CysC (Dy1196 duoset, R&D systems) and MPO (Dy3667 duoset, R&D systems).

Techniques: Biomarker Discovery

Journal: Netherlands Heart Journal

Article Title: The ethnicity-specific association of biomarkers with the angiographic severity of coronary artery disease

doi: 10.1007/s12471-015-0798-y

Figure Lengend Snippet: Regression coefficients (betas with 95 % confidence intervals) of biomarker levels for SYNTAX score

Article Snippet: Commercial antibody combinations were used to quantify NTproBNP (15 C4 and biotinylated 13G12, Hi-test Finland), hsCRP (Dy1707 duoset, R&D systems), CysC (Dy1196 duoset, R&D systems) and MPO (Dy3667 duoset, R&D systems).

Techniques: Biomarker Discovery

Journal: Netherlands Heart Journal

Article Title: The ethnicity-specific association of biomarkers with the angiographic severity of coronary artery disease

doi: 10.1007/s12471-015-0798-y

Figure Lengend Snippet: Results from ROC analysis of biomarker levels for high SYNTAX score (≥ 18 points)

Article Snippet: Commercial antibody combinations were used to quantify NTproBNP (15 C4 and biotinylated 13G12, Hi-test Finland), hsCRP (Dy1707 duoset, R&D systems), CysC (Dy1196 duoset, R&D systems) and MPO (Dy3667 duoset, R&D systems).

Techniques: Biomarker Discovery