primary antibodies against icam1  (ABclonal Biotechnology)

Journal: Journal of Nanobiotechnology

Article Title: New insights into allergic rhinitis treatment: MSC nanovesicles targeting dendritic cells

doi: 10.1186/s12951-024-02748-2

Figure Lengend Snippet: Influence of P-D2-EVs on Fut1-mediated fucosylation of ICAM1. A Western blot analysis of UEA-1-enriched mDCs under different treatments. B Western blot analysis and quantification of α-(1,2)-fucosylation status of ICAM1 protein in each group after UEA-1 enrichment. C IP-based detection and quantification of ICAM1 binding to UEA1 in each group. D Immunofluorescence co-localization (yellow) and quantification of UEA1 and ICAM1, with DAP(I) staining the cell nucleus in blue, scale bar = 25 μm. E Western blot analysis and quantification of α-(1,2)-fucosylation status of ICAM1 protein in each group after UEA-1 enrichment. F IP-based detection and quantification of ICAM1 binding to UEA1 in each group. G Immunofluorescence co-localization (yellow) and quantification of UEA1 and ICAM1, with DAP(I) staining the cell nucleus in blue, scale bar = 25 μm. H Schematic representation of Fut1-OE-iDCs treated with 2DGal; (I) Western blot analysis and quantification of α-(1,2)-fucosylation status of ICAM1 protein in each group after UEA-1 enrichment. J IP-based detection and quantification of ICAM1 binding to UEA1 in each group. K Immunofluorescence co-localization (yellow) and quantification of UEA1 and ICAM1, with DAP(I) staining the cell nucleus in blue, scale bar = 25 μm; UEA1: Ulex europaeus agglutinin 1, 2DGal: 2-deoxy- d -galactose. * indicates statistical significance compared to the control group or between two groups, with P < 0.05; all experiments were repeated three times

Article Snippet: Slides were incubated overnight with primary antibodies against ICAM1 (Thermofisher, MA5-43106) at a dilution of 1:60.

Techniques: Western Blot, Binding Assay, Immunofluorescence, Staining, Control

Journal: Journal of Nanobiotechnology

Article Title: New insights into allergic rhinitis treatment: MSC nanovesicles targeting dendritic cells

doi: 10.1186/s12951-024-02748-2

Figure Lengend Snippet: Influence of P-D2-EVs on DC metabolism via the Fut1/ICAM1/P38 MAPK pathway. A , B Quantification of p-P38 and P38 expression, as well as the p-P38/P38 ratio, in mDCs from different treatment groups using Western blot. C mRNA expression of IL10 in mDCs from different treatment groups measured by RT-qPCR. D Levels of IL10 in the supernatant of mDCs from different treatment groups analyzed using ELISA. E Analysis of intracellular IL-10 levels in mDCs from different treatment groups using flow cytometry. F Schematic representation of the culture conditions for mDCs, mDCs + 2DGal, and m + 2DGal + Anisomycin groups. G Quantification of p-P38 and P38 expression, as well as the p-P38/P38 ratio, in mDCs from different treatment groups using Western blot. H mRNA expression of IL10 in mDCs from different treatment groups measured by RT-qPCR. I Levels of IL10 in the supernatant of mDCs from different treatment groups analyzed using ELISA. J Analysis of intracellular IL-10 levels in mDCs from different treatment groups using flow cytometry; 2DGal: 2-deoxy- d -galactose; *indicates statistical significance compared to the control group or between two groups, with P < 0.05; all experiments were repeated three times

Article Snippet: Slides were incubated overnight with primary antibodies against ICAM1 (Thermofisher, MA5-43106) at a dilution of 1:60.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Control

Journal: Journal of Nanobiotechnology

Article Title: New insights into allergic rhinitis treatment: MSC nanovesicles targeting dendritic cells

doi: 10.1186/s12951-024-02748-2

Figure Lengend Snippet: The impact of P-D2-EVs on regulating the Fut1/ICAM1/P38 MAPK pathway and IL10 metabolism in DCs on Th2 differentiation. A Schematic representation of co-culturing mDCs with CD4 + T cells. B Flow cytometric analysis of the percentage of IL4 and GATA3 in mDCs co-cultured with CD4 + T cells under different treatments. C RT-qPCR analysis of mRNA expression levels of IL4 and GATA3 in mDCs co-cultured with CD4 + T cells under different treatments. D ELISA analysis of IL4 levels in the supernatant of mDCs co-cultured with CD4 + T cells under different treatments. E Flow cytometric analysis of the percentage of IL4 and GATA3 in mDCs co-cultured with CD4 + T cells under different treatments. F RT-qPCR analysis of mRNA expression levels of IL4 and GATA3 in mDCs co-cultured with CD4 + T cells under different treatments. G ELISA analysis of IL4 levels in the supernatant of mDCs co-cultured with CD4 + T cells under different treatments; * indicates statistical significance compared to the control group or between two groups (P < 0.05); all experiments repeated three times. H Schematic diagram of co-culture of CD4 + T cells with mDCs after IL10 receptor blockade. I Flow cytometry analysis of the percentage of intracellular IL4 and GATA3 in CD4 + T cells co-cultured with mDCs under different treatments. J RT-qPCR detection of mRNA expression levels of IL4 and GATA3 in CD4 + T cells co-cultured with mDCs under different treatments. K ELISA analysis of the level of IL4 in the supernatant of co-cultures of mDCs and CD4 + T cells under different treatments. L Schematic diagram of co-culture of CD4 + T cells and mDCs with IL10 antibody. M Flow cytometry analysis of the percentage of intracellular IL4 and GATA3 in CD4 + T cells co-cultured with mDCs and IL10 antibody under different treatments; (N) RT-qPCR detection of mRNA expression levels of IL4 and GATA3 in CD4 + T cells co-cultured with mDCs and IL10 antibody under different treatments. O ELISA analysis of the level of IL4 in the supernatant of co-cultures of mDCs and CD4 + T cells with IL10 antibody. P Schematic diagram of co-culture of CD4 + T cells and mDCs with IL10. Q Flow cytometry analysis of the percentage of intracellular IL4 and GATA3 in CD4 + T cells co-cultured with mDCs and IL10 under different treatments. R RT-qPCR detection of mRNA expression levels of IL4 and GATA3 in CD4 + T cells co-cultured with mDCs and IL10 under different treatments. S ELISA analysis of the level of IL4 in the supernatant of co-cultures of mDCs and CD4 + T cells with IL10; *indicates a significant difference between two groups with P < 0.05, all experiments were repeated three times

Article Snippet: Slides were incubated overnight with primary antibodies against ICAM1 (Thermofisher, MA5-43106) at a dilution of 1:60.

Techniques: Cell Culture, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Control, Co-Culture Assay, Flow Cytometry

Journal: Journal of Nanobiotechnology

Article Title: New insights into allergic rhinitis treatment: MSC nanovesicles targeting dendritic cells

doi: 10.1186/s12951-024-02748-2

Figure Lengend Snippet: In vivo validation of the regulatory mechanism of P-D2-EVs. A Number of nasal itching and sneezing events within 2 h after the final intranasal OVA administration in each group of mice (n = 6). B Total cell count in NALF of each group of mice (n = 6). C OVA-specific serum IgE levels in each group of mice (n = 6). D Representative PAS staining images (scale bar = 100 μm) and quantification of PAS-positive goblet cells in nasal tissue of each group of mice (n = 6). E Representative H&E staining images (scale bar = 100 μm or 25 μm) of nasal tissue, as well as quantification of eosinophils and nasal mucosal thickness, with the red arrow indicating nasal mucosa (n = 6). F Representative images (scale bar = 25 μm) and quantification of neutrophil infiltration in nasal mucosa of each group of mice, with the black arrow indicating positive cells (n = 6). G Protein expression and quantification of Fut1, p-P38, and P3J8 in nasal tissue of each group of mice as detected by Western blotting (n = 6). H α-(1,2)-Fucose glycosylation status and quantification of ICAM1 protein in nasal tissue of each group of mice as detected by Western blotting following UEA-1 enrichment (n = 6). I Binding and quantification of ICAM1 and UEA1 in nasal tissue of each group of mice as detected by co-immunoprecipitation (n = 6). J Expression and quantification of IL10, IL4, IL13, and GATA3 in nasal tissue of each group of mice as detected by RT-qPCR (n = 6). K Levels of IL10, IL4, and IL13 in serum of each group of mice as detected by ELISA (n = 6). L Protein expression and quantification of Fut1, p-P38, and P3J8 in nasal tissue of each group of mice as detected by Western blotting (n = 6). M α-(1,2)-Fucose glycosylation status and quantification of ICAM1 protein in nasal tissue of each group of mice as detected by Western blotting following UEA-1 enrichment (n = 6). N Binding and quantification of ICAM1 and UEA1 in nasal tissue of each group of mice as detected by co-immunoprecipitation (n = 6). O Expression and quantification of IL10, IL4, IL13, and GATA3 in nasal tissue of each group of mice as detected by RT-qPCR (n = 6). P Levels of IL10, IL4, and IL13 in serum of each group of mice as detected by ELISA (n = 6); NALF nasal lavage fluid; *indicates significant difference between two groups with P < 0.05

Article Snippet: Slides were incubated overnight with primary antibodies against ICAM1 (Thermofisher, MA5-43106) at a dilution of 1:60.

Techniques: In Vivo, Cell Counting, Staining, Expressing, Western Blot, Binding Assay, Immunoprecipitation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Fluids and Barriers of the CNS

Article Title: MicroRNA-155 contributes to shear-resistant leukocyte adhesion to human brain endothelium in vitro

doi: 10.1186/s12987-016-0032-3

Figure Lengend Snippet: miR-155 modulates VCAM1 and ICAM1 expression on brain endothelial hCMEC/D3 cells. hCMEC/D3 cell monolayers were transfected with control siRNA ( a , b ) or scrambled Pre-miR and Pre-miR-155 ( c , e ) or Anti-miR and Anti-miR-155 ( d , f ) followed by treatment with a combination of cytokines (TNFα + IFNγ) at 1 ng/ml for 24 h or left unstimulated. a , b Number of shear-resistant firmly adhered Jurkat and THP-1 cells to siVCAM1-hCMEC/D3 monolayer per FOV (640 × 480 μm). c – f VCAM1 and ICAM1 expression levels were quantified by ELISA. Experiments were carried out three and four times with three replicates each. Data are mean ± SEM. Statistical analysis was performed using paired Student’s t test ( *,# P < 0.05, ***,### P < 0.001, # compared to unstimulated)

Article Snippet: Brain endothelial expression of VCAM1 and ICAM1 was measured by cell-surface ELISA performed as previously described [ ] using 2 μg/ml mouse primary antibody against VCAM1 or ICAM1 (R&D SYSTEMS, Abingdon, UK) and the corresponding secondary antibodies conjugated to horseradish peroxidase.

Techniques: Expressing, Transfection, Shear, Enzyme-linked Immunosorbent Assay