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: bioRxiv

Article Title: Selective Immune Silencing by Targeted TGF-β Agonists

doi: 10.64898/2026.01.19.700410

Figure Lengend Snippet: Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.

Article Snippet: The following antibodies were purchased from Miltenyi Biotec: human IgA (130-113-480) and IRF4 (130-100-909).

Techniques: Plasmid Preparation, Construct, SDS Page, Molecular Weight, Recombinant, Flow Cytometry, Expressing, In Vitro, Cell Culture

Journal: bioRxiv

Article Title: Selective Immune Silencing by Targeted TGF-β Agonists

doi: 10.64898/2026.01.19.700410

Figure Lengend Snippet: a. Frequencies of immune cell subsets. b. SCENIC prediction of IRF4 and Blimp1 transcription factor activity. c. Immunoglobulin gene expression. d. UMAP plots showing cell cycle gene signature scores. e. Quantification of cell cycle gene signature scores. f. Scaled signature scores for TGF-β–inducible genes, senescence genes, and apoptosis-related genes, aggregated by sample.

Article Snippet: The following antibodies were purchased from Miltenyi Biotec: human IgA (130-113-480) and IRF4 (130-100-909).

Techniques: Activity Assay, Gene Expression

Journal: BMC Neuroscience

Article Title: Chronic exposure to high fat diet triggers myelin disruption and interleukin-33 upregulation in hypothalamus

doi: 10.1186/s12868-019-0516-6

Figure Lengend Snippet: List of antibodies in the study for immunofluorescence or western blot analysis

Article Snippet: Polyclonal goat anti-IL-33 , R and D Systems Cat#AF3626, RRID:AB_884269 , Escherichia coli -derived recombinant mouse IL-33 , 1:200 (IF).

Techniques: Immunofluorescence, Western Blot, Purification, Affinity Chromatography, Recombinant, Derivative Assay

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