cd69-fitc (cat no. 11-0691, clone h1.2f3)  (Thermo Fisher)

Journal: PLoS ONE

Article Title: Bortezomib Resistance Can Be Reversed by Induced Expression of Plasma Cell Maturation Markers in a Mouse In Vitro Model of Multiple Myeloma

doi: 10.1371/journal.pone.0077608

Figure Lengend Snippet: A. Fluorescence-activated cell sorting analysis of BzS (solid black line), BzR (dark grey histogram) and I-BzR (light grey histogram) cells stained with indicated antibodies. B. Quantitative RT-PCR analysis of Cd93 and Cd69 relative mRNA expression in 595 BzS and BzR and 589 BzS, BzR, and I-BzR cell lines. Values were normalized to Gapd mRNA and error bars represent PCR triplicates. Significance was determined using a one-tailed Student’s t-test (*p<0.05; **p<0.01; ****p<0.0001). C. Fluorescence-activated cell sorting dot plot analysis of BzS and BzR cells double stained with CD93 and CD69 antibodies. Isotype controls for CD69 (FITC) and CD93 (APC) staining have been provided in .

Article Snippet: Cells were stained with the following anti-mouse antibodies: CD184/Cxcr4 PE (clone 2B11, also used for anti-human Cxcr4), CD69 FITC (clone H1.2F3), CD93 APC (clone AA4.1), CD20 PE (clone AISB12), CD22 FITC (clone 2D6), CD27 FITC (clone LG.7F9) (all from eBioscience, San Diego, CA), CD138/syndecan-1 PE (clone 281-2), CD19 PE (clone 1D3), B220 APC (clone RA3-6B2) (all from BD Biosciences, Franklin Lakes, NJ), IgM-FITC (SouthernBiotech, Birmingham, AL), CD38 (BioLegend, San Diego, CA) and analyzed using the FACSCalibur (BD Biosciences, Franklin Lakes, NJ).

Techniques: Fluorescence, FACS, Staining, Quantitative RT-PCR, Expressing, One-tailed Test

Journal: PLoS ONE

Article Title: Bortezomib Resistance Can Be Reversed by Induced Expression of Plasma Cell Maturation Markers in a Mouse In Vitro Model of Multiple Myeloma

doi: 10.1371/journal.pone.0077608

Figure Lengend Snippet: A–B. Fluorescence-activated cell sorting analysis of 589 BzS cells in the presence (+) or absence (−) of 64 nM Bz for 48 hours. Live cells were gated based on FSC and SSC and dot plots representing CD93 and CD69 double stained populations in A. In B., histograms show live untreated BzS cells (solid black line), BzS cells treated with 64 nM Bz for 48 hours (dotted black line) and untreated BzR cells (dark grey histogram) stained with CD93 and CD69 antibodies. C. Quantitative RT-PCR analysis of Cd93 and Cd69 mRNA in 589 lines corresponding to . Values were normalized to Gapd mRNA and error bars represent PCR triplicates. Significance was determined using a one-tailed Student’s t-test (****p<0.0001; NS = not significant).

Article Snippet: Cells were stained with the following anti-mouse antibodies: CD184/Cxcr4 PE (clone 2B11, also used for anti-human Cxcr4), CD69 FITC (clone H1.2F3), CD93 APC (clone AA4.1), CD20 PE (clone AISB12), CD22 FITC (clone 2D6), CD27 FITC (clone LG.7F9) (all from eBioscience, San Diego, CA), CD138/syndecan-1 PE (clone 281-2), CD19 PE (clone 1D3), B220 APC (clone RA3-6B2) (all from BD Biosciences, Franklin Lakes, NJ), IgM-FITC (SouthernBiotech, Birmingham, AL), CD38 (BioLegend, San Diego, CA) and analyzed using the FACSCalibur (BD Biosciences, Franklin Lakes, NJ).

Techniques: Fluorescence, FACS, Staining, Quantitative RT-PCR, One-tailed Test

Journal: PLoS ONE

Article Title: Bortezomib Resistance Can Be Reversed by Induced Expression of Plasma Cell Maturation Markers in a Mouse In Vitro Model of Multiple Myeloma

doi: 10.1371/journal.pone.0077608

Figure Lengend Snippet: A. Quantitative RT-PCR analysis of Irf-4 , Blimp-1 , Ddit3 , and Cxcr4 mRNA expression in 589 untreated cells and following 72 hours of LPS treatment. Values were normalized to Gapd mRNA, and error bars represent PCR triplicates. Significance was determined using a one-tailed Student’s t-test (*p<0.05; **p<0.01; ***p<0.001; ****p<0.0001; NS = not significant). B. Densitometry values representing the percentage of spliced/unspliced (s/u) Xbp-1 in 589 BzS, BzR and I-BzR cells untreated or treated with LPS for 72 hours. C. Quantitative RT-PCR analysis of Cd69 mRNA expression in 589 untreated cells and 72 hour LPS-treated cells. Values were normalized to Gapd mRNA and error bars represent PCR triplicates. Significance was determined using a one-tailed Student’s t-test (*p<0.05; ***p<0.001). D. Fluorescence-activated cell sorting analysis of untreated (solid black line) BzS (top panel), BzR (middle panel), or I-BzR (bottom panel) and LPS-treated (dotted lines) cells stained for CD69 protein expression.

Article Snippet: Cells were stained with the following anti-mouse antibodies: CD184/Cxcr4 PE (clone 2B11, also used for anti-human Cxcr4), CD69 FITC (clone H1.2F3), CD93 APC (clone AA4.1), CD20 PE (clone AISB12), CD22 FITC (clone 2D6), CD27 FITC (clone LG.7F9) (all from eBioscience, San Diego, CA), CD138/syndecan-1 PE (clone 281-2), CD19 PE (clone 1D3), B220 APC (clone RA3-6B2) (all from BD Biosciences, Franklin Lakes, NJ), IgM-FITC (SouthernBiotech, Birmingham, AL), CD38 (BioLegend, San Diego, CA) and analyzed using the FACSCalibur (BD Biosciences, Franklin Lakes, NJ).

Techniques: Quantitative RT-PCR, Expressing, One-tailed Test, Fluorescence, FACS, Staining

Journal: iScience

Article Title: Natural killer cells and dendritic epidermal γδ T cells orchestrate type 1 conventional DC spatiotemporal repositioning toward CD8 + T cells

doi: 10.1016/j.isci.2021.103059

Figure Lengend Snippet: XCR1 promotes NK cell activation and redistribution in the spleen upon MCMV infection (A–C) Spleens of Xcr1 −/− mice and Wt controls were harvested 40 h (A–F), 4 days (G), and 5 days (H) after MCMV infection. Splenic NK cells (NK1.1 + TCRβ − ) were stained for CD69 (A) and intracellularly for IFN-γ (B) and GzmB (C) directly ex vivo . (D) Visualization of activated NK cell clusters in the spleens of MCMV-infected mice. Scale bar: 50 μm. (E) Analysis of NK cell clusters in Xcr1 −/− vs littermate controls upon MCMV infection. Spleen sections were prepared from Karma Cre ;Rosa26 tdRFP ;Xcr1 −/− mice and Karma Cre ;Rosa26 tdRFP ;Xcr1 +/− controls and stained for NKp46 (green), MOMA-1 (purple), IFN-γ (red), and IE-1 (cyan). Inserts show examples of clusters areas as defined in the material and methods. NK cell clusters were identified as groups of at least 10 IFN-γ + cells, encompassing NKp46 + cells and gathered around MCMV-infected cells (IE-1 + ) in the marginal zone (MOMA-1 + ). Scale bar: 50 μm. (F) Quantification of NK cell clusters in the spleens of MCMV-infected Karma Cre ;Rosa26 tdRFP ;Xcr1 −/− mice and their littermate Xcr1 +/− control 40 h after infection. (G) Splenic NK cells (NK1.1 + TCRβ − ) were stained intracellularly for KI67 4 days after infection. (H) Proportion and absolute number of Ly49H + NK cells in Wt vs Xcr1 −/− mice 5 days after infection. One experiment representative of at least 4 independent ones with at least 4 mice per infected group is shown, except for B-E, where two experiments with at least 3 mice per group were pooled. NI, noninfected; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. See also Figure S1 .

Article Snippet: Anti CD69 FITC (Clone H1.2F3) , ThermoFisher eBioscience , #11-0691-82.

Techniques: Activation Assay, Infection, Staining, Ex Vivo, Control

Journal: iScience

Article Title: Natural killer cells and dendritic epidermal γδ T cells orchestrate type 1 conventional DC spatiotemporal repositioning toward CD8 + T cells

doi: 10.1016/j.isci.2021.103059

Figure Lengend Snippet:

Article Snippet: Anti CD69 FITC (Clone H1.2F3) , ThermoFisher eBioscience , #11-0691-82.

Techniques: Purification, Avidin-Biotin Assay, Virus, Recombinant, High Molecular Weight, Cell Isolation, Microarray, Software