cd86 fitc bu63  (Exbio Praha)

Journal: European journal of medical research

Article Title: Olfactory mucosa-mesenchymal stem cells with overexpressed Nrf2 modulate angiogenesis and exert anti-inflammation effect in an in vitro traumatic brain injury model.

doi: 10.1186/s40001-025-02344-6

Figure Lengend Snippet: Fig. 4 Effects of OM-MSCsNrf2 on the microglial polarization in BV2 cells. A–D CD86 MFI (A, B) and CD206 MFI (C, D) in BV2 cells following the co-culture with OM-MSCs (ctrl) and OM-MSCsNrf2. scale bar: 100 μm. E–H Relative mRNA levels of Cd80 (E), Nos2 (F), Arg1 (G), and Fizz1 (H) in BV2 cells following the co-culture with OM-MSCs (ctrl) and OM-MSCsNrf2. Results of independent triplicates were expressed as mean ± standard deviation and the statistical difference between two groups was marked with asterisks (*p < 0.05, **p < 0.01, and ***p < 0.001). OM-MSCs olfactory mucosa-mesenchymal stem cells, Nrf2 Nuclear Factor Erythroid-Derived 2-Like 2, MFI mean fluorescence intensity, Nos2 nitric oxide synthase 2, Arg1: arginase 1

Article Snippet: BV2 cells after the co-culture were fixed in 4% paraformaldehyde (#P1110, Solarbio® Life Sciences, China), treated with 100% pre-chilled methanol (#34885, Sigma, Germany) and incubated with 1% bovine serum albumin (#A8010, Solarbio® Life Sciences, China) for 1 h. Next, the cells were incubated with the following primary antibodies against TMEM119 (label: Alexa Fluor® 647, #ab225494, 1:500, Abcam, UK), CD86 (label: FITC, #NBP2-34569F, 1:500, Novus Biologicals, USA), and CD206 (label: Alexa Fluor® 488, #141709, 1:500, Bio-Legend, Inc., USA) at 4°C overnight.

Techniques: Co-Culture Assay, Standard Deviation, Derivative Assay, Fluorescence

Journal: iScience

Article Title: Reduction of IFN-I responses by plasmacytoid dendritic cells in a longitudinal trans men cohort

doi: 10.1016/j.isci.2023.108209

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Article Snippet: CD86 / FITC / BU63 , BD Biosciences , Cat#: 374204 / RRID: AB_2721574.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software

Journal: iScience

Article Title: Human skin CD141 + dendritic cells regulate cutaneous immunity via the neuropeptide urocortin 2

doi: 10.1016/j.isci.2023.108029

Figure Lengend Snippet: CD141 expressing skin migratory DDCs and VitD3 moDCs share phenotypical and functional characteristics (A) The two major populations of resident DDCs in healthy human skin are CD141 hi CD1c lo (shown in blue); and CD1c hi CD141 – (shown in red). (B) moDCs (shown in black) are largely negative for CD141 while VitD3 treatment upregulates its expression; CD141 lo VitD3 moDCs (shown in orange) and CD141 hi VitD3 moDCs (shown in green). (C‒F) DDC and moDC populations were stained for (C, D) CD83, CD86, CD40, PD-L1, HLA-ABC, HLA-DR, and (E, F) CD14, CD11c, CCR4, CCR6, CCR10, and CLEC9A. (G) moDCs, CD141 lo VitD3 moDCs, and CD141 hi VitD3 moDCs (left panel); CD141 + and CD1c + DDCs (right panel) stimulation of allogeneic CD4 + T cell proliferation, measured ascounts per minute (CPM). (H) CD141 lo VitD3 moDCs and CD141 hi VitD3 moDCs (left panel); CD141 + and CD1c + DDCs (right panel) production of IL-10 in the absence and presence of CD40 ligand-transfected L cells (CD40L Tx). Results are representative of (A, B) more than twenty, (C‒F) three, (G) ten, and (H) five independent experiments. Data are represented as mean ± SEM. Two-way ANOVA (G) or one-way ANOVA (H), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ### p < 0.001. See also Figure S1 .

Article Snippet: Mouse monoclonal anti-human CD86-FITC (Clone: BU63) , Invitrogen , Cat #MA1-10295.

Techniques: Expressing, Functional Assay, Staining, Transfection

Journal: iScience

Article Title: Human skin CD141 + dendritic cells regulate cutaneous immunity via the neuropeptide urocortin 2

doi: 10.1016/j.isci.2023.108029

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Article Snippet: Mouse monoclonal anti-human CD86-FITC (Clone: BU63) , Invitrogen , Cat #MA1-10295.

Techniques: Purification, Recombinant, Staining, Concentration Assay, Sterility, Multiplex Assay, Labeling, Expressing, Transfection, Software

Journal: iScience

Article Title: Myeloid-intrinsic cell cycle-related kinase drives immunosuppression to promote tumorigenesis

doi: 10.1016/j.isci.2023.107626

Figure Lengend Snippet: Ccrk expression maintains the immature status of MDSCs to support tumor growth in vivo (A) Schematic diagram. MDSCs were generated from naive RFP +/+ mouse bone marrows (BMs), treated with IL-6/GM-CSF for four days, and infected with lentivirus (LV)-based sh Ctrl -GFP or sh Ccrk -GFP (1 MOI) for 48 h. The success of virus infection, sorting purifies and expressions of CCRK, p-STAT3 Tyr705 , E4BP4, and ARG-I were confirmed by fluorescent-microscope or western blot, respectively, in FACS sorted RFP + GFP + MDSCs at 48-h post infection. β-actin was served as loading control. 2∗10 6 purified MDSCs were then intratumorally injected into the left or right subcutaneous tumors of the same RIL-175-bearing mouse at day 6 post tumor inoculation for tumor growth analysis. (B) Tumor sizes were measured by caliper every two days and expressed as ½∗(length∗width ). (C and D) (C) Tumor weights, and (D) the absolute numbers of tumor-infiltrating CD8 + T cells were measured at the endpoint (n = 6). (E and F) (E) Blood was collected at 24-h post 2∗10 6 RFP + GFP + MDSCs adoptive transfer in naive C57BL/6 mice for MDSC phenotype analysis in vivo . The representative flow cytometry dot plots and proportions of CD11b + Gr-1 + Ly6G + Ly6C int PMN-MDSCs as well as (F) F4/80 + CD11c - macrophages in RFP + adoptive transferred cells are shown. (G) The expressions of CD86 and CD80 on F4/80 + CD11c - macrophages are shown in overlay histograms and bar charts. (n = 5). Data are presented as mean ± SD. ∗p < 0.05. ∗∗p < 0.01. See also Figures S1 and .

Article Snippet: FITC anti-human CD86 (clone BU63) , Thermo Fisher Scientific , MHCD8601; RRID: AB_10372961.

Techniques: Expressing, In Vivo, Generated, Infection, Virus, Microscopy, Western Blot, Control, Purification, Injection, Adoptive Transfer Assay, Flow Cytometry

Journal: iScience

Article Title: Myeloid-intrinsic cell cycle-related kinase drives immunosuppression to promote tumorigenesis

doi: 10.1016/j.isci.2023.107626

Figure Lengend Snippet: Inactivation of myeloid Ccrk reduces HCC progression by remodeling tumor microenvironment in orthotopic HCC mouse model (A) DNA sequence of Ccrk indel/indel transgenic (TG) mouse. An ATC (encodes amino acid Ile) insertion in Ccrk exon 3 between c.219 and c.220 was generated in C57BL/6 mouse and confirmed by DNA sequencing. (B) The expressions of CCRK, p-STAT3 Tyr705 , STAT3 and E4BP4 in BMs of naive and RIL-175-tumor bearing wild-type (WT) and Ccrk indel/indel mice were detected by western blot. β-actin was served as loading control. (C and D) (C) WT and Ccrk indel/indel mice were inoculated with 5∗10 5 RIL-175 cells via intrahepatic injection and sacrificed at day 21 post tumor inoculation or humane endpoint (n ≥ 9). Liver tumor images and tumor weights at the endpoint are shown, and (D) their survival rates were documented. (E) High-dimensional flow cytometry analysis in tumor-infiltrating CD45 + leukocytes are shown as visualization of clusters in UMAP plots for total and individual groups. UMAP plots shown consists of 40,000 cells each and are representative of concatenated samples within each group. Cell clusters corresponding to CD4 + and CD8 + T cells, NK cells, DCs, macrophages, M-, and PMN-MDSCs are highlighted and their proportions in WT and Ccrk indel/indel mice are shown. (F) Representative overlay histograms and mean fluorescence intensity (MFI) of CD80, CD86, and IL-12 expressions in CD11b − macrophages from WT and Ccrk indel/indel mice are shown. (G and H) (G) Representative flow cytometry dot plots and proportions of IFN-γ + , TNF-ɑ + , IFN-γ + TNF-ɑ + CD4 + T cells or (H) CD8 + T cells in tumor-infiltrating CD45 + leukocytes from WT and Ccrk indel/indel mice are shown. Data are presented as mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01. See also Figures S3 and .

Article Snippet: FITC anti-human CD86 (clone BU63) , Thermo Fisher Scientific , MHCD8601; RRID: AB_10372961.

Techniques: Sequencing, Transgenic Assay, Generated, DNA Sequencing, Western Blot, Control, Injection, Flow Cytometry, Fluorescence

Journal: iScience

Article Title: Myeloid-intrinsic cell cycle-related kinase drives immunosuppression to promote tumorigenesis

doi: 10.1016/j.isci.2023.107626

Figure Lengend Snippet: Knockdown of CCRK reduces the immunosuppressive activity and induces an immune-stimulatory phenotype of MDSCs (A) Schematic diagram. CD33 + MDSCs were generated from healthy donor PBMCs treated with IL-6/GM-CSF for seven days and purified by CD33 microbeads. Purified MDSCs were transfected with siRNA targeting CCRK (si CCRK ) or control (si Ctrl ). Cells were collected and analyzed by qRT-PCR, western blot, or flow cytometry. The antigen-presenting capacity of MDSCs treated with siRNA was determined by mixed leukocyte reaction (MLR) assay via co-culturing them with CD3 microbeads purified and CFSE-labeled CD3 + T cells from a different donor, at a ratio of 1:1 in the presence of human recombinant IL-2 (20 U/mL). T cell proliferation rate and IFN-γ releasing were analyzed by flow cytometry and ELISA, respectively. (B) CCRK expressions in MDSCs treated with si Ctrl or si CCRK are shown at mRNA (relative to GAPDH ) and protein levels. β-actin was served as loading control. Data are presented as mean ± SD from at least five independent experiments. (C) MDSCs labeled with CFSE before siRNA treatment were used to measure MDSC proliferation and CFSE low cells indicated the proliferating population was analyzed by flow cytometry at day three post transfection. (D–F) (D) The expressions of HLA-DR, (E) proportions of CD68 + HLA-DR + macrophages, as well as (F) CD86 and CD80 surface expressions on CD68 + HLA-DR + macrophages were upregulated upon CCRK knockdown in MDSCs. (G and H) (G) MLR assay showed an up-regulated allogenic T cell proliferation indicated by CFSE low proportion and (H) IFN-γ production in T cells co-cultured with MDSC-si CCRK compared to controls. (n = 7). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. See also Figure S5 .

Article Snippet: FITC anti-human CD86 (clone BU63) , Thermo Fisher Scientific , MHCD8601; RRID: AB_10372961.

Techniques: Knockdown, Activity Assay, Generated, Purification, Transfection, Control, Quantitative RT-PCR, Western Blot, Flow Cytometry, Mlr Assay, Labeling, Recombinant, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: iScience

Article Title: Myeloid-intrinsic cell cycle-related kinase drives immunosuppression to promote tumorigenesis

doi: 10.1016/j.isci.2023.107626

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Article Snippet: FITC anti-human CD86 (clone BU63) , Thermo Fisher Scientific , MHCD8601; RRID: AB_10372961.

Techniques: Staining, Virus, Plasmid Preparation, shRNA, Recombinant, Expressing, Transfection, Magnetic Beads, Protein Extraction, Bicinchoninic Acid Protein Assay, Purification, Enzyme-linked Immunosorbent Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: Cell Intrinsic IL-38 Affects B Cell Differentiation and Antibody Production

doi: 10.3390/ijms24065676

Figure Lengend Snippet: Flow cytometry antibodies.

Article Snippet: CD3-PE-CF594 , 145-2C11 , BD , CD86-FITC , BU63 , Immunotools.

Techniques: Flow Cytometry