Journal: Circulation

Article Title: Proteomic Analysis of Lung Tissues From Patients With Pulmonary Arterial Hypertension

doi: 10.1161/circulationaha.110.972745

Figure Lengend Snippet: Figure 4. Western blot analysis of lung homogenates. Equal amounts of protein (20 g) were used for Western blot analysis of PAH (n14) and control (n14) lung homogenates, using antibodies against periostin, CLIC1, CLIC4, haptoglobin, and vinculin. The densi- tometry results (arbitrary units [AU]) of the Western blots are shown as a bar graph. Each bar represents meanSEM. Representative Western blots for each protein are also shown. *P0.05, **P0.01, ***P0.001 vs control.

Article Snippet: Sections were immunostained with an affinity purified rabbit antibody against a recombinant protein epitope (73-217 amino acids) of human CLIC4 (HPA008019; Atlas Antibodies, Stockholm, Sweden; 1-2 μg/ml) and a recombinant human periostin fusion protein (ab14041, Abcam plc; 5 μg/ml).

Techniques: Western Blot, Control

Journal: Circulation

Article Title: Proteomic Analysis of Lung Tissues From Patients With Pulmonary Arterial Hypertension

doi: 10.1161/circulationaha.110.972745

Figure Lengend Snippet: Figure 6. Distribution of CLIC4 immuno- staining in human lung. Sections from lobectomy (A and B) and unused donor lung tissues (C), showing CLIC4 local- ized to bronchiolar epithelium (A), alveo- lar type II cells (open arrows), and mac- rophages (B), and endothelium (arrows) in the pulmonary vasculature (C). Sec- tions of lung from patients with IPAH (D through F), showing CLIC4 and CD31 immunostaining of the endothelium (arrows) in remodeled pulmonary arteries with neointimal proliferation, as well as the distal microvasculature (arrowheads). Cells in the neointimal layer display weaker CLIC4 immunostaining (D and F). Bar50 m.

Article Snippet: Sections were immunostained with an affinity purified rabbit antibody against a recombinant protein epitope (73-217 amino acids) of human CLIC4 (HPA008019; Atlas Antibodies, Stockholm, Sweden; 1-2 μg/ml) and a recombinant human periostin fusion protein (ab14041, Abcam plc; 5 μg/ml).

Techniques: Immunostaining

Journal: Circulation

Article Title: Proteomic Analysis of Lung Tissues From Patients With Pulmonary Arterial Hypertension

doi: 10.1161/circulationaha.110.972745

Figure Lengend Snippet: Figure 7. Distribution of CLIC4 immuno- staining in a plexiform lesion. Adjacent sections of a plexiform lesion, showing the localization of CLIC4 and CD31 in the endothelium (arrows) and CLIC4 in underlying smooth muscle cells or myo- fibroblasts (asterisks). Boxed area (A) shown at higher power (C). Bar50 m.

Article Snippet: Sections were immunostained with an affinity purified rabbit antibody against a recombinant protein epitope (73-217 amino acids) of human CLIC4 (HPA008019; Atlas Antibodies, Stockholm, Sweden; 1-2 μg/ml) and a recombinant human periostin fusion protein (ab14041, Abcam plc; 5 μg/ml).

Techniques: Immunostaining

Journal:

Article Title: Rat CRM1 Is Responsible for the Poor Activity of Human T-Cell Leukemia Virus Type 1 Rex Protein in Rat Cells

doi: 10.1128/JVI.75.23.11515-11525.2001

Figure Lengend Snippet: Effect of overexpressing rat and human CRM1 on Rex activity. (A) REF52 cells were transfected with 0.075 μg of pSRαRex along with 0.5 μg of pDM128RxRE and 0.1 μg of pCDMβ-gal in combination with various amounts of pSRαhCRM1 (⧫) or pSRαrCRM1 (▪). The CAT/β-Gal ratios of all samples were calculated. The ratio of the sample transfected with pSRαRex in the absence of either the hCRM1 or rCRM1 expression plasmid was arbitrarily set to 1. (B) CRM1 proteins produced in REF52 cells that were transfected with 0.3 μg of either rCRM1 or hCRM1 expression plasmids (lane 1, pSRα296; lane 2, pSRαhCRM1; lane 3, pSRαrCRM1) were analyzed by Western blotting as described in Materials and Methods.

Article Snippet: A mouse anti-GAL4 monoclonal antibody (Santa Cruz Biotechnology) and a rabbit anti-Rex C terminus antibody ( 24 ) were used as primary antibodies to detect GAL-fused proteins and the Rex protein, respectively.

Techniques: Activity Assay, Transfection, Expressing, Plasmid Preparation, Produced, Western Blot

Journal:

Article Title: Rat CRM1 Is Responsible for the Poor Activity of Human T-Cell Leukemia Virus Type 1 Rex Protein in Rat Cells

doi: 10.1128/JVI.75.23.11515-11525.2001

Figure Lengend Snippet: Ability of hCRM1 and rCRM1 to support Rex function in HeLa cells. (A) HeLa cells were transfected with 0.05 μg of pSRαRex, 0.5 μg of pDM128RxRE, and 0.1 μg of pCDMβ-gal together with various amounts of pSRαhCRM1 or pSRαrCRM1. At 8 h posttransfection, the medium was replaced with medium containing LMB at 0.6 or 0.8 nM. At 24 h posttransfection, the cells were subjected to CAT and β-Gal measurement. The CAT/β-Gal ratios of all samples were calculated. The ratio of the control sample transfected with pSRαRex in the absence of either hCRM1 or rCRM1 expression plasmids and in the absence of LMB was arbitrarily set to 1. The amounts of CAT and the β-Gal activity in the control sample were over 300 pg and 2.5 × 10−3 U, respectively. (B) CRM1 proteins produced in HeLa cells transfected with 0.3 μg of each CRM1 expression plasmid (lane 1, pSRα296; lane 2, pSRαhCRM1; lane 3, pSRαrCRM1) were analyzed by Western blotting.

Article Snippet: A mouse anti-GAL4 monoclonal antibody (Santa Cruz Biotechnology) and a rabbit anti-Rex C terminus antibody ( 24 ) were used as primary antibodies to detect GAL-fused proteins and the Rex protein, respectively.

Techniques: Transfection, Expressing, Activity Assay, Produced, Plasmid Preparation, Western Blot

Journal:

Article Title: Rat CRM1 Is Responsible for the Poor Activity of Human T-Cell Leukemia Virus Type 1 Rex Protein in Rat Cells

doi: 10.1128/JVI.75.23.11515-11525.2001

Figure Lengend Snippet: Interaction of Rex with either hCRM1 or rCRM1. (A) REF52 cells were transfected with the plasmid expressing either GAL-hCRM1 or GAL-rCRM1 in combination with pRexVP, pG5BLuc, and pCDMβ-gal. The Luc/β-Gal ratios of all samples were calculated. GAL-, plasmid expressing only the GAL4 region, which was used as a negative control. (B) Western blot of GAL-hCRM1 and GAL-rCRM1 proteins synthesized in transfected REF52 cells. (C) Pull-down assay using recombinant Rex protein. Purified His-Rex(g10) proteins immobilized on chelating Sepharose were incubated with HeLa or REF52 cell extract in the absence (lanes 2 and 4) or presence (lanes 3 and 5) of GTP-charged recombinant RanQ69L protein. The sample of lane 1 contained neither cell lysate nor recombinant RanQ69L proteins during incubation. The volume of the cell extract subjected to the binding reaction was nine times of that of the input fraction.

Article Snippet: A mouse anti-GAL4 monoclonal antibody (Santa Cruz Biotechnology) and a rabbit anti-Rex C terminus antibody ( 24 ) were used as primary antibodies to detect GAL-fused proteins and the Rex protein, respectively.

Techniques: Transfection, Plasmid Preparation, Expressing, Negative Control, Western Blot, Synthesized, Pull Down Assay, Recombinant, Purification, Incubation, Binding Assay

Journal:

Article Title: Rat CRM1 Is Responsible for the Poor Activity of Human T-Cell Leukemia Virus Type 1 Rex Protein in Rat Cells

doi: 10.1128/JVI.75.23.11515-11525.2001

Figure Lengend Snippet: Rex-Rex dimerization analyzed by the coimmunoprecipitation method. HeLa and REF52 cells were transfected with the plasmids which express GAL-Rex and Rex-VP or their derivatives as indicated. At 48 h posttransfection, the cells were harvested and the supernatants were incubated with anti-VP antibodies immobilized on protein-G Sepharose in the presence of GTP-charged recombinant RanQ69L. The recombinant hCRM1 protein was added to the sample, which was derived from the REF52 cells transfected with GAL-Rex and Rex-VP (lane 6) during incubation. The volume of the cell extract subjected to the coimmunoprecipitation reaction was nine times of that of input fraction. Rabbit anti-Rex C terminus antibodies were used for Western blotting.

Article Snippet: A mouse anti-GAL4 monoclonal antibody (Santa Cruz Biotechnology) and a rabbit anti-Rex C terminus antibody ( 24 ) were used as primary antibodies to detect GAL-fused proteins and the Rex protein, respectively.

Techniques: Transfection, Incubation, Recombinant, Derivative Assay, Western Blot

Journal: Cell

Article Title: Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease

doi: 10.1016/j.cell.2018.08.067

Figure Lengend Snippet:

Article Snippet: Slides were washed in PBS containing 0.05% Tween 20 (PBS-T) and incubated with ImmPRESS HRP Anti-Rabbit or Anti-Mouse IgG (Peroxidase) Polymer Detection Kit (Vector Laboratories, Peterborough, U.K) for thirty minutes at room temperature.

Techniques: Recombinant, Coagulation, Antibody Labeling, Plasmid Preparation, Blocking Assay, Staining, Imaging, Software, Hybridization

Journal: Bioengineering

Article Title: RNU ( Foxn1 RNU -Nude) Rats Demonstrate an Improved Ability to Regenerate Muscle in a Volumetric Muscle Injury Compared to Sprague Dawley Rats

doi: 10.3390/bioengineering8010012

Figure Lengend Snippet: CD68+ staining is increased in RNU DMM-treated sites compared to Sprague Dawley. CD68 staining was quantified and showed increased levels in RNU rats ( A ). Yet, CD163 ( B ) and FoxP3 ( C ) were unaltered. CD4 staining was increased in RNU rats ( D ) while CD8 staining was lower ( E ). In addition, CD4, CD8, and FoxP3 levels were low. DRAQ5 showed no change in DNA staining ( F ). Data shown are means ± SEM of 8 animals. Letters not shared indicated a significant difference ( p < 0.05, Tukey).

Article Snippet: Primary antibodies used in this experiment were: mouse anti-Pax7 (ab55494, Abcam, Cambridge, UK); rabbit anti-nicotinic acetylcholine receptor-epsilon (AChR-ε, ab65180, Abcam); mouse anti-nicotinic acetylcholine receptor-gamma (AChR-γ, MA3-043, Thermo Fisher Scientific, Waltham, MA, USA); mouse anti-myosin heavy chain-fetal (fMyHC, SC-53097, Santa Cruz Biotechnology); CD68 (ab125212, Abcam); CD163 (ab87099, Abcam); CD8 (MAB116, R&D Systems), CD4 (MAB554, R&D Systems), FoxP3 (MAB8214, R&D Systems) and were diluted in PBS with 1% BSA and 0.3% Tween-20.

Techniques: Staining