Journal: The Journal of Clinical Investigation

Article Title: Innate and adaptive nasal mucosal immune responses following experimental human pneumococcal colonization

doi: 10.1172/JCI128865

Figure Lengend Snippet: ( A ) CyTOF data files were pregated using FlowJo to identify cells (DNA + Bead – ), followed by selecting viable immune cells (CD45 + Dead – ). These cells were exported and loaded in Cytosplore for hierarchical stochastic neighbor embedding (h-SNE), in which lineages, subpopulations, and clusters were sequentially identified in 3 steps. Gating for naive CD4 + T cells is shown as an example. ( B ) Cells were clustered using all 38 markers minus the epithelial marker Epcam and lineages were then defined based on the expression of 9 markers. Clustered lineages and expression of included markers are shown. Subpopulations for ( C ) CD8 + T cells, ( D ) granulocytes, ( E ) CD4 + T cells, ( F ) innate lymphoid cells, ( G ) CD117 + cells, and ( H ) double-negative T cells were defined based on the expression of the depicted markers. B cells, plasma cells, and myeloid cells were not further divided into subpopulations due to lack of clear clustering by relevant markers. Cell subpopulations were then further divided into clusters using all 38 markers minus the epithelial marker Epcam.

Article Snippet: Primary antibodies used were CD3 (Dako), CD4, CD20, CD66b, CD68, CD11b (Abcam), CD8 (Epitomics), and CD161 (Atlas Antibodies), which were applied overnight at 4°C ( ).

Techniques: Marker, Expressing, Clinical Proteomics

Journal: The Journal of Clinical Investigation

Article Title: Innate and adaptive nasal mucosal immune responses following experimental human pneumococcal colonization

doi: 10.1172/JCI128865

Figure Lengend Snippet: To establish an overall cellular distribution in the tissue, an ×10 magnification is shown for each of the markers. Insets (×40 magnification) are also included to visualize some individual positive cells. ( A ) Hematoxylin and eosin (H&E) staining showing the entire biopsy. Staining of subsequent slices showing the biopsy at the epithelial edge for the markers ( B ) negative control, ( C ) CD3, ( D ) CD4, ( E ) CD8, ( F ) CD161, ( G ) CD20, ( H ) CD68, ( I ) CD66b, and ( J ) CD11b. Scale bars: 250 μm. Slices were counterstained with H&E. Some background staining of the extracellular matrix is present for CD161 ( F ). Biopsy was derived from 1 challenged, carriage – subject.

Article Snippet: Primary antibodies used were CD3 (Dako), CD4, CD20, CD66b, CD68, CD11b (Abcam), CD8 (Epitomics), and CD161 (Atlas Antibodies), which were applied overnight at 4°C ( ).

Techniques: Staining, Negative Control, Derivative Assay

Journal: International Journal of Ophthalmology

Article Title: Differentiation of mesenchymal stem cell in the microenviroment of retinitis pigmentosa

doi: 10.3980/j.issn.2222-3959.2010.03.08

Figure Lengend Snippet: A: Brdu(+); B: Rhodopsin(+); C: PCK(+)

Article Snippet: Materials DMEM/F12(1:1) culture medium and fetal calf serum (Hyclone), NaIO 3 , Dulbecco fluid (Gibco), Bromodeoxyuridine (BrdU, Sigma), mouse monoclonal anti-BrdU antibody (Boster Biotechnology Co. Ltd.), PE conjugated anti-rat CD45 monoclonal antibody and FITC conjugated anti-rat CD90 monoclonal antibody (Caltag Laboratory), anti-rat rhodopsin (Opsin) monoclonal antibody (NeoMarkers), anti-rat PCK monoclonal antibody (NeoMarkers), Cy3 conjugated anti-mouse secondary antibody, Sheep blood serum (Boster Biotechnology Co. Ltd.).

Techniques:

Journal: Current protocols in cytometry

Article Title: CyTOF Measurement of Immunocompetence across Major Immune Cell Types

doi: 10.1002/cpcy.27

Figure Lengend Snippet: Example of a Staining Panel

Article Snippet: 145Nd , CD4 , RPA-T4 , Fluidigm.

Techniques: Staining

Journal: The Journal of investigative dermatology

Article Title: TGFbeta1-induced inflammation in premalignant epidermal squamous lesions requires IL-17.

doi: 10.1038/jid.2010.92

Figure Lengend Snippet: Figure 2. TGFb1 induces IL-17 production in CD4 þ and cd þ tumor-infiltrating T cells. Tumor-infiltrating lymphocytes were isolated from papillomas after 2 days of dox treatment and analyzed for cell-surface markers and intracellular cytokines. (a) A representative flow cytometry profile of IL-17 þ and IFNg þ T cells from ST and DT papillomas. (b) Percentage of IL-17 þ and IFNg þ CD4 þ T cells from ST (n ¼ 11) and DT (n ¼ 12) papillomas, and IL-17 þ

Article Snippet: For immunohistochemistry, anti-human MPO (Dako, Carpinteria, CA); anti-CD3e (Santa Cruz Biotechnology, Santa Cruz, CA); CD4 and CD8 (Angio-Proteomie, Boston, MA); and anti-phospho Smad2 (Santa Cruz Biotechnology) antibodies were used on 70% ethanol- fixed sections after antigen retrieval.

Techniques: Isolation, Flow Cytometry

Journal: The Journal of investigative dermatology

Article Title: TGFbeta1-induced inflammation in premalignant epidermal squamous lesions requires IL-17.

doi: 10.1038/jid.2010.92

Figure Lengend Snippet: Figure 4. TGFb1 overexpression increases inflammatory infiltrate in benign tumors. ST papillomas (a, c) and DT papillomas (b, d–f) were harvested from mice placed on dox for 1 week. Tumors were stained with hematoxylin and eosin-Y (a, b), antiMPO for myeloid cells (c, d), and anti-CD3e (e) and anti-CD8 (f). Bars ¼ 20 mM. The arrows indicate infiltrating leukocytes. The photomicrographs are representative of at least three different tumors per genotype. S, stroma; T, tumor. (g–i) Quantitation of tumor-infiltrating T cells from immunohistochemical analysis for CD3 þ (g), CD4 þ (h), and CD8 þ (i) cells. No staining was detected in the absence of primary antibody. Each histogram represents the average from at least four tumors of each genotype, with more than five fields counted per section. Error bars ¼ ±SEM.

Article Snippet: For immunohistochemistry, anti-human MPO (Dako, Carpinteria, CA); anti-CD3e (Santa Cruz Biotechnology, Santa Cruz, CA); CD4 and CD8 (Angio-Proteomie, Boston, MA); and anti-phospho Smad2 (Santa Cruz Biotechnology) antibodies were used on 70% ethanol- fixed sections after antigen retrieval.

Techniques: Over Expression, Staining, Quantitation Assay, Immunohistochemical staining

Journal: Molecular medicine reports

Article Title: microRNAs mediate oleic acid-induced acute lung injury in rats using an alternative injury mechanism.

doi: 10.3892/mmr.2014.2155

Figure Lengend Snippet: Figure 4. IHC with monoclonal anti-α-actin antibody was performed 6 and 24 h after OA-induced ALI. The figures are representative microphotographs from each category. (A) IHC of a control group sample at 6 h. Positivity for α-SMA is observed in tangentially sectioned basal epithelial cells and along the BM of bronchial epithelium; (B) IHC of the control group 24 h after OA-induced ALI. Fluorescent activity in the alveolar wall was similar to the IHC result at 6 h; (C) IHC of the OA group sample 6 h after OA-induced ALI. Alveolar wall edema and mucus secretion in the alveolar space were observed (asterisk). DAB staining in the epithelial cells (arrowhead) and in a wide area of stromal connective tissue underneath the BM was observed. (D) Increased DAB staining in the alveolar BM (arrowhead), smooth muscle of the bronchial wall or of the blood vessels (arrows) was revealed 24 h after OA-induced ALI. Thick mucus secretion was found in the alveolar spaces (asterisk) 24 h after OA-induced ALI. IHC, immunohistochemical staining; OA, oleic acid; ALI, acute lung injury; BM, basement membrane; DAB, diaminobenzidine.

Article Snippet: The measurement of the concentration of IL-13 in bronchoalveolar lavage (BAL) supernatant was performed by ELISA with rat monoclonal primary antibodies and goat polyclonal secondary antibodies, according to the manufacturer's instructions (Cusabio Biotech Co., Ltd., Wuhan, Hubei, China; Cat no. 84290).

Techniques: Control, Activity Assay, Staining, Immunohistochemical staining, Membrane