staining with anti cd3 fitc  (Elabscience Biotechnology)

Journal: Animals : an Open Access Journal from MDPI

Article Title: Silica–Calcite Sedimentary Rock (Opoka) Enhances the Immunological Status and Improves the Growth Rate in Broilers Exposed to Ochratoxin A in Feed

doi: 10.3390/ani14010024

Figure Lengend Snippet: Lymphocyte, leukocyte, and monocyte levels in the blood of broiler chickens fed with control feed with ochratoxin and feed with ochratoxin and Opoka. Different letters signify statistically significant group differences ( p < 0.05).

Article Snippet: Specific monoclonal antibodies, including mouse anti-chicken CD3 + and CD4 + (fluorescein isothiocyanate, FITC staining), CD8 + , Bu-1 + and monocytic/macrophagic + antibodies (phycoerythrin, PE staining) (SouthernBiotech, Birmingham, AL, USA), were then employed.

Techniques: Control

Journal: bioRxiv

Article Title: Epigenetic compound screening uncovers small molecules for re-activation of latent HIV-1

doi: 10.1101/2020.08.21.262311

Figure Lengend Snippet: Resting CD4 + T-cells were isolated from blood of healthy donors and challenged for 24 h with either PHA or CD3/CD28 beads (positive controls) or with the respective compound. Vehicle-treated cells were used as negative control (control). Subsequently, cells were stained for CD3, CD4, CD25 and CD69 and analyzed by flow cytometry for increased expression of activation markers CD25 and CD69. Flow cytometry plots show gating for CD3+/CD4+ subset (left) and corresponding flow cytometry plot (right) shows distribution of CD25/CD69 on control cells (black) and activated, PHA-treated cells (red). Bar charts indicate amount of CD25, CD69, and CD25/CD69 positive cells. Error bars represent SD form three different donors.

Article Snippet: Subsequently, activation of T-cells was measured by flow cytometry using the anti-human CD4 (FITC)/CD69 (PE)/CD3 (PerCP) antibody staining solution (BD Biosciences) in combination with anti CD25 (APC) (BD Biosciences).

Techniques: Isolation, Negative Control, Staining, Flow Cytometry, Expressing, Activation Assay

Journal: bioRxiv

Article Title: Epigenetic compound screening uncovers small molecules for re-activation of latent HIV-1

doi: 10.1101/2020.08.21.262311

Figure Lengend Snippet: PBMCs, derived from healthy donors, were challenged for 24 h with the respective compound or with vehicle control and subsequently analyzed by flow cytometry for certain cellular markers. The flow cytometry plots show representative data of control cells and the underlying gating strategy. Cells were first gated for singlets, tested for viability via 7-AAD stain, and cell debris was removed by classical SSC/FSC analysis. Derived living single cells were analyzed for CD45 expression as general leucocyte marker and leucocytes were further discriminated in B-cells (CD19) and monocytes (CD14). CD19/CD14 negative cells were than subdivided into eosinophils/granulocytes (SSC high) and CD16dim SSC low fraction. CD3 and CD56 were used for determination of CD3+ T-cells, CD56+ NK cells and CD3+/CD56+ T-cell like NK-cells. Subsequently, CD3+ cells were analyzed for CD4+ and CD8+ T-cells. Associated data for each cell type is presented as bar chart diagram and shows results from three different donors. Error bars represent SD.

Article Snippet: Subsequently, activation of T-cells was measured by flow cytometry using the anti-human CD4 (FITC)/CD69 (PE)/CD3 (PerCP) antibody staining solution (BD Biosciences) in combination with anti CD25 (APC) (BD Biosciences).

Techniques: Derivative Assay, Flow Cytometry, Staining, Expressing, Marker

Journal: Nature Communications

Article Title: Immunoprophylactic and immunotherapeutic control of hormone receptor-positive breast cancer

doi: 10.1038/s41467-020-17644-0

Figure Lengend Snippet: a Schedule of oncogenic challenges for the induction of M/D-driven tumors and representative images of M/D-driven tumors established in WT C57BL/6 mice. Scale bar = 1 cm. b Tumor-free survival (TFS) and time-to-death (TTD) in WT C57BL/6 mice subjected to M/D-driven oncogenesis. Number of mice is reported. c Representative histology of M/D-driven tumors as compared to human HR + breast cancers. Scale bar = 50 µm. d Probeset distribution comparing the transcriptomic profile of 6 M/D-driven mammary tumors established in C57BL/6 mice with that of human breast cancers from the TCGA public database. Probability of molecular subtyping is reported for each mouse tumors. e , f TFS and overall survival (OS) of WT C57BL/6 mice and C57BL/6 bearing ER mutations that cause nuclear exclusion ( e ) or nuclear accumulation ( f ) subjected to M/D-driven oncogenesis. Number of mice, hazard ratio (HR) and p values (two-sided log-rank) are reported. g TFS and OS of WT C57BL/6 mice subjected to M/D-driven oncogenesis in control conditions or along with tamoxifen administration in the drinking water. Number of mice, HR and p values (two-sided log-rank) are reported. h . Non-supervised hierarchical clustering of the transcriptomic profile of M/D-driven tumors established in WT C57BL/6 mice ( n = 6), mammary glands exposed to M/D but not developing tumors ( n = 6) and M/D-naïve mammary glands ( n = 6). Gene Ontology analysis, fold change (FC) and adjusted, two-sided p values are reported. Red, upregulation. Yellow, downregulation. i Relative amount of CD3 + , CD8 + , CD4 + and CD25 + FOXP3 + cells infiltrating M/D-driven tumors in C57BL/6 mice vs syngeneic M/D-naïve mammary glands. Results are means ± SEM plus individual data points. Number of mice and p values (unpaired, two-sided Student’s t , as compared to M/D-naïve mammary glands) are reported.

Article Snippet: Gating procedure is exemplified in Supplementary Fig. . Alternatively, cells were stained with LIVE/DEAD™ Fixable Yellow dye and Fc receptors were blocked as described above, then fixed in Cytofix/Cytoperm™ buffer after surface staining with anti-CD3-FITC (1:800, clone 17A2, #11-0032-82 from Thermo Fisher), anti-NK1.1-PerCP-Cy5.5 (1:100, clone PK136, #551114 from BD Biosciences), anti-B220-V450 (1:100, clone RA3-6B2, # 560473 from BD Biosciences) and anti-CD19-APC-Vio770 (1:50, clone REA749, #130-111-886 from Miltenyi).

Techniques: