Journal: Science Advances

Article Title: The tumor suppressor adenomatous polyposis coli regulates T lymphocyte migration

doi: 10.1126/sciadv.abl5942

Figure Lengend Snippet: Primary and secondary antibodies/reagents used for immunofluorescence.

Article Snippet: APC anti-human CD11a , Mouse IgG1 , Miltenyi Biotec 130-127-294 , 1:50.

Techniques: Immunofluorescence, Purification

Journal: Science Advances

Article Title: The tumor suppressor adenomatous polyposis coli regulates T lymphocyte migration

doi: 10.1126/sciadv.abl5942

Figure Lengend Snippet: Antibodies used for flow cytometry.

Article Snippet: APC anti-human CD11a , Mouse IgG1 , Miltenyi Biotec 130-127-294 , 1:50.

Techniques: Cytometry

Journal: Science Advances

Article Title: The tumor suppressor adenomatous polyposis coli regulates T lymphocyte migration

doi: 10.1126/sciadv.abl5942

Figure Lengend Snippet: Antibodies used for Western blot.

Article Snippet: APC anti-human CD11a , Mouse IgG1 , Miltenyi Biotec 130-127-294 , 1:50.

Techniques: Western Blot

Journal: Diabetes

Article Title: Cardioprotective and anti-inflammatory effects of interleukin converting enzyme inhibition in experimental diabetic cardiomyopathy.

doi: 10.2337/db06-1662

Figure Lengend Snippet: FIG. 3. Representative histological pictures of CD68 and CD11b cells (magnification 200 and 1,000, marked by black arrows) of an STZ animal. CD11b cell with 1,000 magnification demonstrating an intravascular leukocyte before undergoing transendothelial migration. Bars representing data from histological evaluation of immunocompetent cells positive for the macrophage marker CD68 and for the leukocyte markers CD18, CD11a, and CD11b in cardiac tissue of control, STZ-induced diabetes, and STZ treated with the ICEI (STZICEI). *Significantly different compared with control and STZICEI, with P < 0.05. §Significantly different compared with control and STZ.

Article Snippet: Staining was performed with the following primary antibodies at the dilutions given (45 min, room temperature): for mouse anti-rat: ICAM-1 (clone 1A29, 1:100; Serotec, Munich, Germany), VCAM-1 (clone 5F10, 1:50; BabCO, Richmond, VA), CD18 (clone WT3, 1:50; Serotec), CD11a (clone WT1, 1:100; Serotec), CD11b (clone MRC OX-42, 1:100; Biozol, Eching, Germany), COL 1 (1:200; Abcam Limited, Cambridge, U.K.), Col3 (clone FH-7A, 1:40.000; Abcam Limited), and CD68 (clone ED1, 1:2,000; Serotec); for goat anti-rat: TNF- (1:25; R&D Systems, Wiesbaden, Germany) and IL1- (1:25; R&D Systems); and for rabbit anti-rat: IL-18 (1:25; R&D Systems).

Techniques: Migration, Marker, Control

Journal: Cell Reports Medicine

Article Title: Response and recurrence correlates in individuals treated with neoadjuvant anti-PD-1 therapy for resectable oral cavity squamous cell carcinoma

doi: 10.1016/j.xcrm.2021.100411

Figure Lengend Snippet:

Article Snippet: CD11a (HI111) , Fluidigm , Cat# 3142006B, RRID: AB_2877095.

Techniques: Recombinant, Isolation, Mass Cytometry, Software

Journal: Infection and Immunity

Article Title: Aggregatibacter actinomycetemcomitans Leukotoxin (LtxA) Requires Death Receptor Fas, in Addition to LFA-1, To Trigger Cell Death in T Lymphocytes

doi: 10.1128/IAI.00309-19

Figure Lengend Snippet: Generation and validation of CD11a knockout in Jurkat cells. (A) DNA from the independent clones with mutations in CD11a was isolated, PCR amplified, cloned into a TOPO vector, and sequenced. Sequences were aligned to wild-type Jurkat E6.1 CD11a DNA. Any alteration in the DNA sequence was considered gene editing. Each dash indicates a single nucleotide deletion; lowercase letters indicate nucleotide insertions. The number of nucleotides inserted or deleted is indicated at the end of each sequence. (B) Flow cytometric analysis of Jurkat E6.1 cells and both Jurkat CD11a−/− clones for surface expression of CD11a. Cell populations are indicated according to the legend on the figure. (C) Western blot analysis to confirm deletion of CD11a and the GAPDH loading control. Gels were spliced for labeling purposes.

Article Snippet: Anti-human CD11a (clone MAB3595; R&D Systems) (1:500 dilution), anti-human CD18 (D4N5Z; Cell Signaling Technology) (1:1,000 dilution), anti-human Fas (clone 4C3; Cell Signaling Technology) (1:1,000 dilution) and anti-glyceraldehyde-3-phosphate dehydrogenase conjugated to horseradish peroxidase (GAPDH-HRP; 1:2,500 dilution) (clone 1E6D9; ProteinTech) were used as primary antibodies for Western blot analyses.

Techniques: Biomarker Discovery, Knock-Out, Clone Assay, Isolation, Amplification, Plasmid Preparation, Sequencing, Expressing, Western Blot, Control, Labeling

Journal: Infection and Immunity

Article Title: Aggregatibacter actinomycetemcomitans Leukotoxin (LtxA) Requires Death Receptor Fas, in Addition to LFA-1, To Trigger Cell Death in T Lymphocytes

doi: 10.1128/IAI.00309-19

Figure Lengend Snippet: CD11a is required for LtxA-mediated cell death. (A) Jurkat E6.1 cells and both Jurkat CD11a−/− clones were treated with LtxA for 24 h. Staurosporine (STS; 1 μM) was used as a positive control for cell death. (B and C) Jurkat E6.1 cells and Jurkat CD11a−/− clones were treated with 0.1 μg/ml or 1.0 μg/ml LtxA for 24, 48, or 72 h, and cell death was assessed via flow cytometry. Cytotoxicity was determined by flow cytometry, and the percent cell death is defined as the sum of annexin V+/7-AAD− and annexin V+/7-AAD+ cells. Data represent the average from three independent experiments. Error bars represent SEM. The significance of differences between results for Jurkat E6.1 cells and those for each Jurkat CD11a−/− clone was determined using Student's t test. ***, P ≤ 0.001; ns, not significant.

Article Snippet: Anti-human CD11a (clone MAB3595; R&D Systems) (1:500 dilution), anti-human CD18 (D4N5Z; Cell Signaling Technology) (1:1,000 dilution), anti-human Fas (clone 4C3; Cell Signaling Technology) (1:1,000 dilution) and anti-glyceraldehyde-3-phosphate dehydrogenase conjugated to horseradish peroxidase (GAPDH-HRP; 1:2,500 dilution) (clone 1E6D9; ProteinTech) were used as primary antibodies for Western blot analyses.

Techniques: Clone Assay, Positive Control, Flow Cytometry

Journal: Infection and Immunity

Article Title: Aggregatibacter actinomycetemcomitans Leukotoxin (LtxA) Requires Death Receptor Fas, in Addition to LFA-1, To Trigger Cell Death in T Lymphocytes

doi: 10.1128/IAI.00309-19

Figure Lengend Snippet: CD18 and CD11a are required for LtxA to activate caspases. (A) Jurkat E6.1 cells and Jurkat CD18−/− clones were treated with various concentrations of LtxA for 24 h. Caspase activation was assessed using a fluorescent polycaspase reagent and flow cytometry. Increases in fluorescent signal are directly related to the amount of active caspases within a cell. Representative images are shown. (B) Cells were gated on buffer-treated controls, and fold change in caspase activation was determined using FlowJo. (C) Jurkat E6.1 cells and both Jurkat CD11a−/− clones were treated with various concentrations of LtxA for 24 h, and caspase activation was assessed via flow cytometry. Representative images are shown. (D) Cells were gated on buffer controls and fold change in caspase activation was determined using FlowJo. Cell populations are indicated according to the legend on the figure. Data represent the average of at least three independent experiments. Error bars represent SEM. The significance of differences between results for Jurkat E6.1 cells and those of Jurkat CD18−/− clones or Jurkat CD11a−/− clones was determined using Student's t test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.

Article Snippet: Anti-human CD11a (clone MAB3595; R&D Systems) (1:500 dilution), anti-human CD18 (D4N5Z; Cell Signaling Technology) (1:1,000 dilution), anti-human Fas (clone 4C3; Cell Signaling Technology) (1:1,000 dilution) and anti-glyceraldehyde-3-phosphate dehydrogenase conjugated to horseradish peroxidase (GAPDH-HRP; 1:2,500 dilution) (clone 1E6D9; ProteinTech) were used as primary antibodies for Western blot analyses.

Techniques: Clone Assay, Activation Assay, Flow Cytometry

Journal: Infection and Immunity

Article Title: Aggregatibacter actinomycetemcomitans Leukotoxin (LtxA) Requires Death Receptor Fas, in Addition to LFA-1, To Trigger Cell Death in T Lymphocytes

doi: 10.1128/IAI.00309-19

Figure Lengend Snippet: Characterization of Jurkat Fas knockout cells. (A) DNA from the independent clones with mutations in Fas was isolated, PCR amplified, cloned into a TOPO vector, and sequenced. Sequences were aligned to wild-type Jurkat E6.1 Fas DNA. Any alteration in the DNA sequence was considered gene editing. Each lowercase letter indicates a nucleotide insertion; the number of nucleotides inserted is indicated at the end of each sequence. Sequence of the clone with a deletion in Fas exon 4 (top) and of the clone with a deletion in Fas exon 6 (bottom). (B) Flow cytometric analysis of Jurkat E6.1 cells and both Jurkat Fas−/− clones for surface expression of Fas. Cell populations are indicated according to the legend on the figure. (C) Western blot analysis to confirm deletion of Fas and GAPDH as a loading control. Gels were spliced for labeling purposes. (D and E) Flow cytometric analysis for surface expression of CD11a (D) and CD18 (E) in Jurkat Fas−/− clones. Cell populations are indicated according to the legend on the figure. (F) Jurkat E6.1 cells and Jurkat Fas−/− clones were treated with 10 ng/ml FasL for 24 h, and cell death was assessed via flow cytometry and annexin V/7-AAD staining. Data represent the average of three independent experiments. Error bars represent SEM. The significance of differences between results for Jurkat E6.1 cells and those for each Jurkat Fas−/− clone was determined using Student's t test. ***, P ≤ 0.001.

Article Snippet: Anti-human CD11a (clone MAB3595; R&D Systems) (1:500 dilution), anti-human CD18 (D4N5Z; Cell Signaling Technology) (1:1,000 dilution), anti-human Fas (clone 4C3; Cell Signaling Technology) (1:1,000 dilution) and anti-glyceraldehyde-3-phosphate dehydrogenase conjugated to horseradish peroxidase (GAPDH-HRP; 1:2,500 dilution) (clone 1E6D9; ProteinTech) were used as primary antibodies for Western blot analyses.

Techniques: Knock-Out, Clone Assay, Isolation, Amplification, Plasmid Preparation, Sequencing, Expressing, Western Blot, Control, Labeling, Flow Cytometry, Staining

Journal: Veterinary Research

Article Title: Porcine CD18 mediates Actinobacillus pleuropneumoniae ApxIII species-specific toxicity

doi: 10.1051/vetres/2009016

Figure Lengend Snippet: Susceptibility to ApxIIIA-induced cell death closely depends on the rate of LFA-1 ectopic expression in human erythroleukemic K-562 cells provided the contributing CD18 subunit is that of the pig. Values (squares, circles, triangles and lozenges) are means ± SD from three representative experiments in which mean LFA-1 expression rate (horizontal SD) and mean PI incorporation (vertical SD) were measured by flow cytometry. LFA-1 cell-surface expression rate typical of each experiment was defined as the mean proportion of Alexa 488-positive cells calculated from two independent labelling procedures, one using MCA1972 (anti-CD18) and the other MCA2308 or 555382 (anti-CD11a) as primary antibody.

Article Snippet: MAbs MCA1972 (anti-pig CD18) and MCA2308 (anti-pig CD11a) were purchased from Abd Serotec (Düsseldorf, Germany), mAb BAQ30A (anti-bovine CD18) from VMRD (Pullman, USA), mAb 555382 (anti-human/bovine CD11a) from BD Biosciences (Erembodegem, Belgium) and AlexaFluor ® 488-conjugated goat anti-mouse IgG from Invitrogen (Carlsbad, CA, USA).

Techniques: Expressing, Flow Cytometry