pe cyanine7 anti mouse cd80  (Elabscience Biotechnology)

Journal: Stem Cell Research & Therapy

Article Title: Migrasomes derived from human umbilical cord mesenchymal stem cells: a new therapeutic agent for ovalbumin-induced asthma in mice

doi: 10.1186/s13287-025-04145-4

Figure Lengend Snippet: Administration of hUCMSC-migrasomes diminished Th2 response in allergic asthma by suppressing DC maturation. A The expression of IL-4, IL-5 and IL-13 in the lung of mice in each group were detected by real-time PCR ( n = 6). B Gating strategy and representative flow cytometric images for the analysis of CD45 + CD4 + CCR4 + CXCR3 − Th2 cells in the lungs of mice from each group. C Graphs showing the % of CD45 + CD4 + CCR4 + CXCR3 − Th2 cells in the lungs of mice from each group ( n = 6). D Gating strategy and representative flow cytometric dot plots (left) for the analysis of CD11b + DCs in the lungs of allergic mice and the numbers of CD11b + DCs (right) in the lungs of mice from each group ( n = 6). E The mean MFI of CD80, CD86 and MHC-II on CD11b + DCs ( n = 4–6). F The expression of IL-6 in the lung of mice were detected by real-time PCR ( n = 6). G Distribution of Dir-labeled migrasomes in OVA-induced mice lung after tail vein administration (17-day injection and 18-day detected). H Representative immunofluorescence images of Dil-labeled migrasomes (red) taken up by CD11c + cells or CD4 + cells. Data were presented as mean ± SD. A one-way analysis of variance (Tukey Kramer post hoc tests) was performed on the data. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The cell pellets were then stained with a range of monoclonal antibodies including FITC anti-mouse CD45 (eBioscience, San Diego, CA), eflour 450 anti-mouse CD4 (eBioscience), PE- anti-mouse CCR4 (Biolegend), APC anti-mouse CXCR3 (Biolegend), eflour 450 anti-mouse CD11c (eBioscience), FITC anti-mouse CD11b (Elabscience, Wuhan, China), PE/Cyanine7 anti-mouse CD80 (Elabscience), PE anti-mouse CD86 (eBioscience) and APC anti-mouse MHC-II (eBioscience).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Labeling, Injection, Immunofluorescence

Journal: Stem Cell Research & Therapy

Article Title: Migrasomes derived from human umbilical cord mesenchymal stem cells: a new therapeutic agent for ovalbumin-induced asthma in mice

doi: 10.1186/s13287-025-04145-4

Figure Lengend Snippet: HUCMSC-migrasomes impaired DC maturation and function in vitro. A BMDCs incubated with Dil-labeled migrasomes under fluorescence microscope (white bar = 25 μm). B The cell viability of BMDCs treated by hUCMSC-migrasomes for 24 h (left) and 48 h (right) ( n = 4). C BMDCs were isolated from wild-type BALB/c mice and cultured at a density of 2 × 10⁵ cells/mL. The cells were stimulated with OVA (100 µg/mL) and LPS (10 ng/mL) for 48 h, with or without hUCMSC-migrasomes (20 µg/mL) pre-treated. After that, cells were collected for the detection of DC maturation markers including CD80, CD86 and MHC-II ( n = 3). The representative images for the detection of DC maturation markers are displayed. D Statistical analysis of the mean MFIs of CD80, CD86 and MHC-II of BMDCs ( n = 3). E Statistical analysis of the expression of IL-6 in BMDCs by real-time PCR ( n = 3). F BMDCs (2 × 10 4 ) from WT mice were pre-treated with or without migrasomes and then pulsed with OVA 323–339 and LPS and incubated at a 1:5 ratio with splenic CD4 + T cells isolated from OT-II mice (1 × 10 5 ) for 72 h. G Statistical analysis of the expression of IL-4, IL-5, IL-13 in collected cell by real-time PCR ( n = 3). Data were presented as mean ± SD. A one-way analysis of variance (Tukey Kramer post hoc tests) was performed on the data. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The cell pellets were then stained with a range of monoclonal antibodies including FITC anti-mouse CD45 (eBioscience, San Diego, CA), eflour 450 anti-mouse CD4 (eBioscience), PE- anti-mouse CCR4 (Biolegend), APC anti-mouse CXCR3 (Biolegend), eflour 450 anti-mouse CD11c (eBioscience), FITC anti-mouse CD11b (Elabscience, Wuhan, China), PE/Cyanine7 anti-mouse CD80 (Elabscience), PE anti-mouse CD86 (eBioscience) and APC anti-mouse MHC-II (eBioscience).

Techniques: In Vitro, Incubation, Labeling, Fluorescence, Microscopy, Isolation, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Interrelationship of Stress, Environment, and Herpes Simplex Virus Type-1 on Behçet’s Disease: Using a Mouse Model

doi: 10.3389/fimmu.2021.607768

Figure Lengend Snippet: Frequencies of DC costimulatory molecules CD40, CD83, CD80, and CD86 in peripheral blood leukocytes (PBLs) were evaluated in normal, HSV-1 inoculated, BDN, and BD mice. Isolated PBLs were analyzed by flow cytometry after surface staining (A–D) . Representative histogram of CD83+ cells is shown in (E) . For statistical analysis, the Mann-Whitney U test was performed with GraphPad. Experiments were performed independently at least three times.

Article Snippet: Then 1x10 6 cells were stained with Percp-eFluor-labeled anti-mouse CD40, eFluor 660-labeled anti-mouse CD83, PE-Cyanine7-labeled anti-mouse CD80, and FITC-labeled anti-mouse CD86 (eBioscience, San Diego, CA, USA) at 4°C in the dark for 30 min. Stained cells were analyzed with FACS Aria III flow cytometer (Becton Dickinson, San Jose, CA, USA) with x10,000 gated cells.

Techniques: Isolation, Flow Cytometry, Staining, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Interrelationship of Stress, Environment, and Herpes Simplex Virus Type-1 on Behçet’s Disease: Using a Mouse Model

doi: 10.3389/fimmu.2021.607768

Figure Lengend Snippet: Expression levels of dendritic cell activation markers in normal mice after stress treatment. Frequencies of DC activation molecules CD40+, CD83+, CD80+, and CD86+ cells in normal mice after stress treatment were analyzed by FACS analysis (A–D) . Substance P (SP) was used for anxiety stress, 4°C temperature was used for cold stress, xanthine sodium salt was used for oxidative stress, and 77dB sound was used for noise stress induction. For statistical analysis, Mann-Whitney U test was performed. Cellular transcription factor mRNA expression levels in PBLs of normal mice exposed to noise stress were analyzed by real time-PCR (E–H) .

Article Snippet: Then 1x10 6 cells were stained with Percp-eFluor-labeled anti-mouse CD40, eFluor 660-labeled anti-mouse CD83, PE-Cyanine7-labeled anti-mouse CD80, and FITC-labeled anti-mouse CD86 (eBioscience, San Diego, CA, USA) at 4°C in the dark for 30 min. Stained cells were analyzed with FACS Aria III flow cytometer (Becton Dickinson, San Jose, CA, USA) with x10,000 gated cells.

Techniques: Expressing, Activation Assay, MANN-WHITNEY, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Interrelationship of Stress, Environment, and Herpes Simplex Virus Type-1 on Behçet’s Disease: Using a Mouse Model

doi: 10.3389/fimmu.2021.607768

Figure Lengend Snippet: Frequency analysis of DC-stimulating molecules CD40, CD83, CD80 and CD86-expressing cells in normal mice and HSV-1 inoculated mice with or without noise stress. Mice were exposed to noise stress for 10 days after the first HSV-1 inoculation. One day after second inoculation of HSV-1, peripheral blood leukocytes were applied to flow cytometry (A–D) . Experiments were conducted independently at least three times.

Article Snippet: Then 1x10 6 cells were stained with Percp-eFluor-labeled anti-mouse CD40, eFluor 660-labeled anti-mouse CD83, PE-Cyanine7-labeled anti-mouse CD80, and FITC-labeled anti-mouse CD86 (eBioscience, San Diego, CA, USA) at 4°C in the dark for 30 min. Stained cells were analyzed with FACS Aria III flow cytometer (Becton Dickinson, San Jose, CA, USA) with x10,000 gated cells.

Techniques: Expressing, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Interrelationship of Stress, Environment, and Herpes Simplex Virus Type-1 on Behçet’s Disease: Using a Mouse Model

doi: 10.3389/fimmu.2021.607768

Figure Lengend Snippet: Noise stress affects DC activation in BD mice. To investigate the correlation between noise stress and DC activation in BD, frequencies of DC activation markers CD40, CD83, CD80, and CD83 positive cells in peripheral blood leukocytes of noise-treated BD mice and BDN mouse were measured by flow cytometry (A–D) . BD mice under noise stress showed deteriorated or new symptoms (E) . Experiments were conducted independently at least three times. mRNA expression levels of cell transcription factors ROR-γt and Gata-3 in mice with only ocular symptoms and BD mice with ocular symptoms were analyzed by real time PCR. Both groups of mice were compared with mice in noise stress treated groups (F, G) . Statistical analysis was performed with Student’s t -test. The experiment was conducted in duplicate.

Article Snippet: Then 1x10 6 cells were stained with Percp-eFluor-labeled anti-mouse CD40, eFluor 660-labeled anti-mouse CD83, PE-Cyanine7-labeled anti-mouse CD80, and FITC-labeled anti-mouse CD86 (eBioscience, San Diego, CA, USA) at 4°C in the dark for 30 min. Stained cells were analyzed with FACS Aria III flow cytometer (Becton Dickinson, San Jose, CA, USA) with x10,000 gated cells.

Techniques: Activation Assay, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Interrelationship of Stress, Environment, and Herpes Simplex Virus Type-1 on Behçet’s Disease: Using a Mouse Model

doi: 10.3389/fimmu.2021.607768

Figure Lengend Snippet: CD83 siRNA reduces frequencies of CD83+ cells in BD mice. Both noise stress and CD83 inhibition were used to treat BD mice at the same time. Symptoms were improved and frequencies of CD83+ cells were decreased in the group that showed inhibition of CD83 after treatment with CD83 siRNA. BD mice were treated with noise stress and CD83 inhibition at the same time and changes in symptoms and frequencies of DC activation markers were analyzed (A–D) . In the group that showed inhibition of CD83 after treatment with CD83 siRNA, frequencies of CD83+ cells were decreased (B) . CD83 siRNA inhibited the increase in frequencies of CD83+ cells induced by noise stress and inhibited the deterioration of BD symptoms (E) . For statistical analysis, Mann-Whitney U test was performed. Experiments were conducted independently at least three times.

Article Snippet: Then 1x10 6 cells were stained with Percp-eFluor-labeled anti-mouse CD40, eFluor 660-labeled anti-mouse CD83, PE-Cyanine7-labeled anti-mouse CD80, and FITC-labeled anti-mouse CD86 (eBioscience, San Diego, CA, USA) at 4°C in the dark for 30 min. Stained cells were analyzed with FACS Aria III flow cytometer (Becton Dickinson, San Jose, CA, USA) with x10,000 gated cells.

Techniques: Inhibition, Activation Assay, MANN-WHITNEY