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apc anti il 1β  (Miltenyi Biotec)


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    Miltenyi Biotec apc anti il 1β
    Apc Anti Il 1β, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/apc+anti+il+1%CE%B2/IL-1%CE%B2+Antibody%2C+anti-human%2C+REAfinity/pmc10562697-240-42-46
    Average 93 stars, based on 10 article reviews
    apc anti il 1β - by Bioz Stars, 2026-09
    93/100 stars

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    Staining:

    Article Title: Targeting the chemokine receptor CXCR4 with histamine analogue to reduce inflammation in juvenile arthritis: a proof of concept for COVID-19 therapeutic approach
    Article Snippet: For the TNF-α and IL-1β intracellular staining, an Inside Stain kit (Miltenyi Biotec) was used according to the manufacturer’s protocol. .. Briefly, the cells were fixed for 20 min at RT with 250 μL of the Inside Fix solution then washed and stained in 100 μL of the Inside Perm solution containing PE anti-TNF (clone cA2, Miltenyi Biotec) and APC anti-IL-1β (clone REA1172, Miltenyi Biotec) antibody at 1/50 for 30 min at room temperature. .. Data acquisition was performed on a Canto II flow cytometer using Diva software (BD Biosciences, San Jose, CA).

    Article Title: Targeting the chemokine receptor CXCR4 with histamine analog to reduce inflammation in juvenile arthritis
    Article Snippet: For intracellular staining of TNF-α and IL-1β, an Inside Stain kit (Miltenyi Biotec) was utilized in accordance with the manufacturer’s instructions. .. In brief, the cells were fixed with 250 μL of the Inside Fix solution for 20 minutes at room temperature (RT), then washed and stained with 100 μL of the Inside Perm solution containing PE anti TNF-α (clone cA2, Miltenyi Biotec) and APC anti-IL-1β (clone REA1172, Miltenyi Biotec) antibodies at a dilution of 1/50 for 30 minutes at RT. .. Data acquisition was carried out using a Canto II flow cytometer and analyzed using Diva software (BD Biosciences, San Jose, CA) and FlowJo software (Treestar, Ashland, OR).



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    Clinical Characteristics of Gout Patients and Healthy Controls

    Journal: Journal of Inflammation Research

    Article Title: Role of Chemotaxis of Vδ2 T Cells to the Synovium in the Pathogenesis of Acute Gouty Arthritis

    doi: 10.2147/JIR.S449329

    Figure Lengend Snippet: Clinical Characteristics of Gout Patients and Healthy Controls

    Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated anti-human CD14, phycoerythrin (PE)-conjugated anti-human CD16, allophycocyanine (APC)-conjugated anti-human CD192 (CCR2), PE-conjugated anti-human CD294 (CRTH2), APC-conjugated anti-human CD127 (IL-7Rα), peridinin-chlorophyll-protein/cyanine5.5 (PerCP/Cy5.5) – conjugated anti-human CD117 (c-kit), FITC-conjugated anti-human Lineage Cocktail (CD3, CD14, CD19, CD20, CD56), PerCP/Cy5.5-conjugated anti-human CD3, APC-conjugated anti-human CD57, PerCP-conjugated anti-human CD8, PE/Cy7-conjugated anti-human CD56 (NCAM), FITC-conjugated anti-human CD3, PE-conjugated anti-human TCR γ/δ, APC anti-human TCR Vδ2, PerCP/Cy5.5-conjugated anti-human CD3, FITC-conjugated anti-human TCR Vδ2, APC-conjugated anti-human IL-1β, APC-conjugated anti-human IL-17, APC-conjugated anti-human tumor necrosis factor (TNF) -α, APC-conjugated anti-human interferon (IFN) -γ, PE/Cy7-conjugated anti-human C-X-C chemokine receptor 3 (CXCR3), and APC/Fire 750-conjugated anti-human C-C chemokine receptor 5 (CCR5) antibodies were purchased from BD Biosciences (Franklin Lakes, NJ, USA).

    Techniques:

    Patients with AGA had low levels of peripheral Vδ2 T cells. ( A ) Illustrated in this figure are representative scatter plots delineating the gating strategy employed for the detection of γδ T cells, Vδ1 T cells, and Vδ2 T cells. Lymphocytes are identified by their forward scatter (FSC) and side scatter (SSC) characteristics, with single cells isolated based on side scatter area (SSC-A) and side scatter height (SSC-H) parameters. Subsequent gating is conducted for γδ T cells using CD3 and TCR γδ markers, and for Vδ1 and Vδ2 T cells using Vδ1 and Vδ2 markers, respectively. ( B-C ) Peripheral blood mononuclear cells derived from HCs and patients with AGA were subjected to staining with anti-CD3, anti-γδ TCR, anti-Vδ1 or anti-Vδ2 monoclonal antibodies (mAbs) for subsequent flow cytometry analysis. The left panels show representative flow cytometry profiles of γδ T cells ( B ), and Vδ1 T cells and Vδ2 T cells ( C ). The right panels show bar graphs of the percentage of the respective cell populations. The sample size consists of n=18 for the HC group, and n=27 for the AGA group. ( D ) The percentage of peripheral Vδ2 T cells in AGA shows a negative correlation with the concentration of CRP, value of ESR and the count of neutrophils (n=21). ( E-G ) Depicted herein are alternations in the percentages of γδ T cells, Vδ2 T cells and Vδ1 T cells across HCs and different stages of gout. Data from HC (n=18), acute period (n=27), remission period (n=11) and intercritical period (n=26) are shown. Results are expressed as mean±SEM. ns, no significance; * P <0.05, and *** P <0.001 signify significance as determined through two-tailed unpaired t -test. Correlations are calculated using Spearman correlation analysis.

    Journal: Journal of Inflammation Research

    Article Title: Role of Chemotaxis of Vδ2 T Cells to the Synovium in the Pathogenesis of Acute Gouty Arthritis

    doi: 10.2147/JIR.S449329

    Figure Lengend Snippet: Patients with AGA had low levels of peripheral Vδ2 T cells. ( A ) Illustrated in this figure are representative scatter plots delineating the gating strategy employed for the detection of γδ T cells, Vδ1 T cells, and Vδ2 T cells. Lymphocytes are identified by their forward scatter (FSC) and side scatter (SSC) characteristics, with single cells isolated based on side scatter area (SSC-A) and side scatter height (SSC-H) parameters. Subsequent gating is conducted for γδ T cells using CD3 and TCR γδ markers, and for Vδ1 and Vδ2 T cells using Vδ1 and Vδ2 markers, respectively. ( B-C ) Peripheral blood mononuclear cells derived from HCs and patients with AGA were subjected to staining with anti-CD3, anti-γδ TCR, anti-Vδ1 or anti-Vδ2 monoclonal antibodies (mAbs) for subsequent flow cytometry analysis. The left panels show representative flow cytometry profiles of γδ T cells ( B ), and Vδ1 T cells and Vδ2 T cells ( C ). The right panels show bar graphs of the percentage of the respective cell populations. The sample size consists of n=18 for the HC group, and n=27 for the AGA group. ( D ) The percentage of peripheral Vδ2 T cells in AGA shows a negative correlation with the concentration of CRP, value of ESR and the count of neutrophils (n=21). ( E-G ) Depicted herein are alternations in the percentages of γδ T cells, Vδ2 T cells and Vδ1 T cells across HCs and different stages of gout. Data from HC (n=18), acute period (n=27), remission period (n=11) and intercritical period (n=26) are shown. Results are expressed as mean±SEM. ns, no significance; * P <0.05, and *** P <0.001 signify significance as determined through two-tailed unpaired t -test. Correlations are calculated using Spearman correlation analysis.

    Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated anti-human CD14, phycoerythrin (PE)-conjugated anti-human CD16, allophycocyanine (APC)-conjugated anti-human CD192 (CCR2), PE-conjugated anti-human CD294 (CRTH2), APC-conjugated anti-human CD127 (IL-7Rα), peridinin-chlorophyll-protein/cyanine5.5 (PerCP/Cy5.5) – conjugated anti-human CD117 (c-kit), FITC-conjugated anti-human Lineage Cocktail (CD3, CD14, CD19, CD20, CD56), PerCP/Cy5.5-conjugated anti-human CD3, APC-conjugated anti-human CD57, PerCP-conjugated anti-human CD8, PE/Cy7-conjugated anti-human CD56 (NCAM), FITC-conjugated anti-human CD3, PE-conjugated anti-human TCR γ/δ, APC anti-human TCR Vδ2, PerCP/Cy5.5-conjugated anti-human CD3, FITC-conjugated anti-human TCR Vδ2, APC-conjugated anti-human IL-1β, APC-conjugated anti-human IL-17, APC-conjugated anti-human tumor necrosis factor (TNF) -α, APC-conjugated anti-human interferon (IFN) -γ, PE/Cy7-conjugated anti-human C-X-C chemokine receptor 3 (CXCR3), and APC/Fire 750-conjugated anti-human C-C chemokine receptor 5 (CCR5) antibodies were purchased from BD Biosciences (Franklin Lakes, NJ, USA).

    Techniques: Isolation, Derivative Assay, Staining, Flow Cytometry, Concentration Assay, Two Tailed Test

    The Proportions of γδT Cells and Their Subtypes in Gout Patients and Healthy Controls

    Journal: Journal of Inflammation Research

    Article Title: Role of Chemotaxis of Vδ2 T Cells to the Synovium in the Pathogenesis of Acute Gouty Arthritis

    doi: 10.2147/JIR.S449329

    Figure Lengend Snippet: The Proportions of γδT Cells and Their Subtypes in Gout Patients and Healthy Controls

    Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated anti-human CD14, phycoerythrin (PE)-conjugated anti-human CD16, allophycocyanine (APC)-conjugated anti-human CD192 (CCR2), PE-conjugated anti-human CD294 (CRTH2), APC-conjugated anti-human CD127 (IL-7Rα), peridinin-chlorophyll-protein/cyanine5.5 (PerCP/Cy5.5) – conjugated anti-human CD117 (c-kit), FITC-conjugated anti-human Lineage Cocktail (CD3, CD14, CD19, CD20, CD56), PerCP/Cy5.5-conjugated anti-human CD3, APC-conjugated anti-human CD57, PerCP-conjugated anti-human CD8, PE/Cy7-conjugated anti-human CD56 (NCAM), FITC-conjugated anti-human CD3, PE-conjugated anti-human TCR γ/δ, APC anti-human TCR Vδ2, PerCP/Cy5.5-conjugated anti-human CD3, FITC-conjugated anti-human TCR Vδ2, APC-conjugated anti-human IL-1β, APC-conjugated anti-human IL-17, APC-conjugated anti-human tumor necrosis factor (TNF) -α, APC-conjugated anti-human interferon (IFN) -γ, PE/Cy7-conjugated anti-human C-X-C chemokine receptor 3 (CXCR3), and APC/Fire 750-conjugated anti-human C-C chemokine receptor 5 (CCR5) antibodies were purchased from BD Biosciences (Franklin Lakes, NJ, USA).

    Techniques:

    Vδ2 T cells accumulated in gout synovial fluid. ( A and B ) The proportions of γδ T cells within the lymphocytes subset ( A ) and the ratios of Vδ2 T cells and Vδ1 T cells among the total γδ T cells populations ( B ) were quantified through flow cytometry analyses for OA-SFMC samples (n=9) and AGA-SFMC samples (n=9). Representative flow profiles are shown and the percentages were summarized. ( C and D ) The left panels show representative flow profiles of γδ T cells ( C ), Vδ1 T cells and Vδ2 T cells ( D ) derived from both PBMC, and SFMC from patients with gout (n=8). The right panels show bar graphs of the percentage of corresponding cells within the indicated populations. Results are expressed as mean±SEM. ns, no significance; ** P <0.01, and *** P <0.001 signify significance as determined through two-tailed paired t -test.

    Journal: Journal of Inflammation Research

    Article Title: Role of Chemotaxis of Vδ2 T Cells to the Synovium in the Pathogenesis of Acute Gouty Arthritis

    doi: 10.2147/JIR.S449329

    Figure Lengend Snippet: Vδ2 T cells accumulated in gout synovial fluid. ( A and B ) The proportions of γδ T cells within the lymphocytes subset ( A ) and the ratios of Vδ2 T cells and Vδ1 T cells among the total γδ T cells populations ( B ) were quantified through flow cytometry analyses for OA-SFMC samples (n=9) and AGA-SFMC samples (n=9). Representative flow profiles are shown and the percentages were summarized. ( C and D ) The left panels show representative flow profiles of γδ T cells ( C ), Vδ1 T cells and Vδ2 T cells ( D ) derived from both PBMC, and SFMC from patients with gout (n=8). The right panels show bar graphs of the percentage of corresponding cells within the indicated populations. Results are expressed as mean±SEM. ns, no significance; ** P <0.01, and *** P <0.001 signify significance as determined through two-tailed paired t -test.

    Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated anti-human CD14, phycoerythrin (PE)-conjugated anti-human CD16, allophycocyanine (APC)-conjugated anti-human CD192 (CCR2), PE-conjugated anti-human CD294 (CRTH2), APC-conjugated anti-human CD127 (IL-7Rα), peridinin-chlorophyll-protein/cyanine5.5 (PerCP/Cy5.5) – conjugated anti-human CD117 (c-kit), FITC-conjugated anti-human Lineage Cocktail (CD3, CD14, CD19, CD20, CD56), PerCP/Cy5.5-conjugated anti-human CD3, APC-conjugated anti-human CD57, PerCP-conjugated anti-human CD8, PE/Cy7-conjugated anti-human CD56 (NCAM), FITC-conjugated anti-human CD3, PE-conjugated anti-human TCR γ/δ, APC anti-human TCR Vδ2, PerCP/Cy5.5-conjugated anti-human CD3, FITC-conjugated anti-human TCR Vδ2, APC-conjugated anti-human IL-1β, APC-conjugated anti-human IL-17, APC-conjugated anti-human tumor necrosis factor (TNF) -α, APC-conjugated anti-human interferon (IFN) -γ, PE/Cy7-conjugated anti-human C-X-C chemokine receptor 3 (CXCR3), and APC/Fire 750-conjugated anti-human C-C chemokine receptor 5 (CCR5) antibodies were purchased from BD Biosciences (Franklin Lakes, NJ, USA).

    Techniques: Flow Cytometry, Derivative Assay, Two Tailed Test

    CXCR3 and CXCL10 may be associated with the chemotaxis of Vδ2 T cell in AGA. ( A ) Transwell migration assay demonstrated a pronounced capacity of chemotaxis in Vδ2 T cells isolated from AGA patients. Data were pooled from three independent experiments. ( B ) Comparison of the proportion of CXCR3-positive cells in Vδ2 T cells from HCs and AGA patients. ( C ) Comparison of the proportion of CXCR3-positive cells in Vδ2 T cells and Vδ1 T cells from AGA patients. ( D ) Comparison of the proportion of CCR5-positive cells in Vδ2 T cells from HCs and AGA patients. Data of HC (n=8) and AGA (n=10) are shown. ( E and F ) The concentrations of established ligands of CXCR3 and CCR5 were quantified within SF of gout patients (n=18), RA patients (n=7), OA patients (n=7), and serum of gout patients (n=49). Results are expressed as mean±SEM. * P <0.05, ** P <0.01, and *** P <0.001 signify significance as determined through the Mann–Whitney U -test ( A and E ) and two-tailed unpaired t -test ( B–D ).

    Journal: Journal of Inflammation Research

    Article Title: Role of Chemotaxis of Vδ2 T Cells to the Synovium in the Pathogenesis of Acute Gouty Arthritis

    doi: 10.2147/JIR.S449329

    Figure Lengend Snippet: CXCR3 and CXCL10 may be associated with the chemotaxis of Vδ2 T cell in AGA. ( A ) Transwell migration assay demonstrated a pronounced capacity of chemotaxis in Vδ2 T cells isolated from AGA patients. Data were pooled from three independent experiments. ( B ) Comparison of the proportion of CXCR3-positive cells in Vδ2 T cells from HCs and AGA patients. ( C ) Comparison of the proportion of CXCR3-positive cells in Vδ2 T cells and Vδ1 T cells from AGA patients. ( D ) Comparison of the proportion of CCR5-positive cells in Vδ2 T cells from HCs and AGA patients. Data of HC (n=8) and AGA (n=10) are shown. ( E and F ) The concentrations of established ligands of CXCR3 and CCR5 were quantified within SF of gout patients (n=18), RA patients (n=7), OA patients (n=7), and serum of gout patients (n=49). Results are expressed as mean±SEM. * P <0.05, ** P <0.01, and *** P <0.001 signify significance as determined through the Mann–Whitney U -test ( A and E ) and two-tailed unpaired t -test ( B–D ).

    Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated anti-human CD14, phycoerythrin (PE)-conjugated anti-human CD16, allophycocyanine (APC)-conjugated anti-human CD192 (CCR2), PE-conjugated anti-human CD294 (CRTH2), APC-conjugated anti-human CD127 (IL-7Rα), peridinin-chlorophyll-protein/cyanine5.5 (PerCP/Cy5.5) – conjugated anti-human CD117 (c-kit), FITC-conjugated anti-human Lineage Cocktail (CD3, CD14, CD19, CD20, CD56), PerCP/Cy5.5-conjugated anti-human CD3, APC-conjugated anti-human CD57, PerCP-conjugated anti-human CD8, PE/Cy7-conjugated anti-human CD56 (NCAM), FITC-conjugated anti-human CD3, PE-conjugated anti-human TCR γ/δ, APC anti-human TCR Vδ2, PerCP/Cy5.5-conjugated anti-human CD3, FITC-conjugated anti-human TCR Vδ2, APC-conjugated anti-human IL-1β, APC-conjugated anti-human IL-17, APC-conjugated anti-human tumor necrosis factor (TNF) -α, APC-conjugated anti-human interferon (IFN) -γ, PE/Cy7-conjugated anti-human C-X-C chemokine receptor 3 (CXCR3), and APC/Fire 750-conjugated anti-human C-C chemokine receptor 5 (CCR5) antibodies were purchased from BD Biosciences (Franklin Lakes, NJ, USA).

    Techniques: Chemotaxis Assay, Transwell Migration Assay, Isolation, Comparison, MANN-WHITNEY, Two Tailed Test

    Vδ2 T cells in patients with AGA secreted high levels of IL-17. ( A ) Flow cytometry analyses were conducted to assess intracellular IL-17 staining in Vδ2 T cells sourced from peripheral blood samples of both HCs and AGA patients. The right panels show the percentage of positively stained cells among Vδ2 T cells. Data of HCs (n=7) and AGA (n=9) are shown. ( B ) Statistical graph showed the levels of various types of cytokines in gout SF (n=6), serum from patients with AGA (n=7) and serum from HCs (n=7). ( C and D ) Flow cytometry analyses of the intracellular staining of IL-17 in Vδ2 T cells ( C ) and in Th17 cells ( D ) of PBMC and SFMC from patients with gout were performed. The right panels show the percentage of positively stained cells among the indicated populations. Data of both groups (n=8) are shown. Results are expressed as mean±SEM. ns, no significance; * P <0.05, ** P <0.01, and *** P <0.001 signify significance as determined through by two-tailed unpaired t -test.

    Journal: Journal of Inflammation Research

    Article Title: Role of Chemotaxis of Vδ2 T Cells to the Synovium in the Pathogenesis of Acute Gouty Arthritis

    doi: 10.2147/JIR.S449329

    Figure Lengend Snippet: Vδ2 T cells in patients with AGA secreted high levels of IL-17. ( A ) Flow cytometry analyses were conducted to assess intracellular IL-17 staining in Vδ2 T cells sourced from peripheral blood samples of both HCs and AGA patients. The right panels show the percentage of positively stained cells among Vδ2 T cells. Data of HCs (n=7) and AGA (n=9) are shown. ( B ) Statistical graph showed the levels of various types of cytokines in gout SF (n=6), serum from patients with AGA (n=7) and serum from HCs (n=7). ( C and D ) Flow cytometry analyses of the intracellular staining of IL-17 in Vδ2 T cells ( C ) and in Th17 cells ( D ) of PBMC and SFMC from patients with gout were performed. The right panels show the percentage of positively stained cells among the indicated populations. Data of both groups (n=8) are shown. Results are expressed as mean±SEM. ns, no significance; * P <0.05, ** P <0.01, and *** P <0.001 signify significance as determined through by two-tailed unpaired t -test.

    Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated anti-human CD14, phycoerythrin (PE)-conjugated anti-human CD16, allophycocyanine (APC)-conjugated anti-human CD192 (CCR2), PE-conjugated anti-human CD294 (CRTH2), APC-conjugated anti-human CD127 (IL-7Rα), peridinin-chlorophyll-protein/cyanine5.5 (PerCP/Cy5.5) – conjugated anti-human CD117 (c-kit), FITC-conjugated anti-human Lineage Cocktail (CD3, CD14, CD19, CD20, CD56), PerCP/Cy5.5-conjugated anti-human CD3, APC-conjugated anti-human CD57, PerCP-conjugated anti-human CD8, PE/Cy7-conjugated anti-human CD56 (NCAM), FITC-conjugated anti-human CD3, PE-conjugated anti-human TCR γ/δ, APC anti-human TCR Vδ2, PerCP/Cy5.5-conjugated anti-human CD3, FITC-conjugated anti-human TCR Vδ2, APC-conjugated anti-human IL-1β, APC-conjugated anti-human IL-17, APC-conjugated anti-human tumor necrosis factor (TNF) -α, APC-conjugated anti-human interferon (IFN) -γ, PE/Cy7-conjugated anti-human C-X-C chemokine receptor 3 (CXCR3), and APC/Fire 750-conjugated anti-human C-C chemokine receptor 5 (CCR5) antibodies were purchased from BD Biosciences (Franklin Lakes, NJ, USA).

    Techniques: Flow Cytometry, Staining, Two Tailed Test

    Properdin knockout ameliorates microglial activation and inflammation in tMCAO mice. a qPCR assessment of differentially expressed inflammatory genes, including Il1a , Il1b , Il4 , Il6 , Il10 , Nos2 , Ccl2 and Tnf, in the ischemic penumbra 1 d and 3 d after tMCAO. n = 5 mice per group. * P < 0.05 versus the sham group, ** P < 0.01 versus the sham group, # P < 0.05 versus the WT group, ## P < 0.01 versus the WT group, one-way ANOVA with Bonferroni post hoc test. b Immunostaining for TMEM119 (green) and Iba1 (red) and 3D-reconstructed images of TMEM119 + Iba1 + microglia 1 d and 3 d after tMCAO. Scale bar: 50 µm. n = 5 mice per group. * P < 0.05 versus the WT group, ** P < 0.01 versus the WT group, unpaired Student’s t test. c Quantification of Iba1 MFI. d Quantification of average branch length, branch numbers and junctions of 3D-reconstructed microglia in the peri-infarct region. e One day after tMCAO, microglia sorted based on the CD45 int and CD11b + strategy were subjected to PCR array analysis. f FACS analysis of IL-1β, TNF-α and IL-6 expression in microglia 1 d after tMCAO. MFI was quantified. n = 5 mice per group. *** P < 0.001 versus the sham group, ## P < 0.01 versus the WT group, one-way ANOVA with Bonferroni post hoc test

    Journal: Journal of Neuroinflammation

    Article Title: Infiltrating myeloid cell-derived properdin markedly promotes microglia-mediated neuroinflammation after ischemic stroke

    doi: 10.1186/s12974-023-02946-z

    Figure Lengend Snippet: Properdin knockout ameliorates microglial activation and inflammation in tMCAO mice. a qPCR assessment of differentially expressed inflammatory genes, including Il1a , Il1b , Il4 , Il6 , Il10 , Nos2 , Ccl2 and Tnf, in the ischemic penumbra 1 d and 3 d after tMCAO. n = 5 mice per group. * P < 0.05 versus the sham group, ** P < 0.01 versus the sham group, # P < 0.05 versus the WT group, ## P < 0.01 versus the WT group, one-way ANOVA with Bonferroni post hoc test. b Immunostaining for TMEM119 (green) and Iba1 (red) and 3D-reconstructed images of TMEM119 + Iba1 + microglia 1 d and 3 d after tMCAO. Scale bar: 50 µm. n = 5 mice per group. * P < 0.05 versus the WT group, ** P < 0.01 versus the WT group, unpaired Student’s t test. c Quantification of Iba1 MFI. d Quantification of average branch length, branch numbers and junctions of 3D-reconstructed microglia in the peri-infarct region. e One day after tMCAO, microglia sorted based on the CD45 int and CD11b + strategy were subjected to PCR array analysis. f FACS analysis of IL-1β, TNF-α and IL-6 expression in microglia 1 d after tMCAO. MFI was quantified. n = 5 mice per group. *** P < 0.001 versus the sham group, ## P < 0.01 versus the WT group, one-way ANOVA with Bonferroni post hoc test

    Article Snippet: To evaluate the levels of inflammatory factors in microglia in vivo and in vitro, the cells were further incubated with the following antibodies: PE-anti-TNF-α (Rat, 1:200, Thermo Fisher Scientific, OH, USA, Cat# 12-7321-81, RRID:AB_466198), PE-anti-IL-6 (Rat, 1:200, BioLegend, CA, USA, Cat# 504503, RRID:AB_315337) and APC-anti-IL-1β. (Rabbit, 1:500, Cell Signaling Technology, MA, USA, Cat# 31202, RRID:AB_2799001) All the samples were analyzed by a BD LSRFortessa flow cytometer (BD, NJ, USA), and the data were further analyzed using FlowJo software (Ashland, OR, USA, RRID:SCR_008520).

    Techniques: Knock-Out, Activation Assay, Immunostaining, Expressing

    rmProperdin activates microglia and induces microglia-potentiated neuronal death in vitro. a The mRNA expression of inflammatory factors was measured in primary microglia treated with 2 µg/ml, 4 µg/ml and 8 µg/ml rmProperdin versus control microglia. n = 3 per group, * P < 0.05 versus the control group, ** P < 0.01 versus the control group, *** P < 0.001 versus the control group, n.s, no significance ( P > 0.05), one-way ANOVA with Bonferroni post hoc test. b FACS analysis of IL-1β, TNF-α and IL-6 expression in primary microglia treated with rmProperdin for 24 h. MFI was quantified. n = 3 per group, ** P < 0.01 versus the control group, *** P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test. c Experimental design for the analysis of neuronal viability after treatment with rmProperdin or CM from rmProperdin-treated microglia. d Neuronal viability was assessed by CCK8 assay. n = 6 per group, *** P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test. e Analysis of neuronal death using calcein-AM (green)/PI (red) double staining. Scale bar: 50 µm. f Calcein-AM-positive primary cortical neurons were quantified as percentages of total cells. n = 5 per group, ** P < 0.01 versus the control group, *** P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test

    Journal: Journal of Neuroinflammation

    Article Title: Infiltrating myeloid cell-derived properdin markedly promotes microglia-mediated neuroinflammation after ischemic stroke

    doi: 10.1186/s12974-023-02946-z

    Figure Lengend Snippet: rmProperdin activates microglia and induces microglia-potentiated neuronal death in vitro. a The mRNA expression of inflammatory factors was measured in primary microglia treated with 2 µg/ml, 4 µg/ml and 8 µg/ml rmProperdin versus control microglia. n = 3 per group, * P < 0.05 versus the control group, ** P < 0.01 versus the control group, *** P < 0.001 versus the control group, n.s, no significance ( P > 0.05), one-way ANOVA with Bonferroni post hoc test. b FACS analysis of IL-1β, TNF-α and IL-6 expression in primary microglia treated with rmProperdin for 24 h. MFI was quantified. n = 3 per group, ** P < 0.01 versus the control group, *** P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test. c Experimental design for the analysis of neuronal viability after treatment with rmProperdin or CM from rmProperdin-treated microglia. d Neuronal viability was assessed by CCK8 assay. n = 6 per group, *** P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test. e Analysis of neuronal death using calcein-AM (green)/PI (red) double staining. Scale bar: 50 µm. f Calcein-AM-positive primary cortical neurons were quantified as percentages of total cells. n = 5 per group, ** P < 0.01 versus the control group, *** P < 0.001 versus the control group, one-way ANOVA with Bonferroni post hoc test

    Article Snippet: To evaluate the levels of inflammatory factors in microglia in vivo and in vitro, the cells were further incubated with the following antibodies: PE-anti-TNF-α (Rat, 1:200, Thermo Fisher Scientific, OH, USA, Cat# 12-7321-81, RRID:AB_466198), PE-anti-IL-6 (Rat, 1:200, BioLegend, CA, USA, Cat# 504503, RRID:AB_315337) and APC-anti-IL-1β. (Rabbit, 1:500, Cell Signaling Technology, MA, USA, Cat# 31202, RRID:AB_2799001) All the samples were analyzed by a BD LSRFortessa flow cytometer (BD, NJ, USA), and the data were further analyzed using FlowJo software (Ashland, OR, USA, RRID:SCR_008520).

    Techniques: In Vitro, Expressing, Control, CCK-8 Assay, Double Staining