primary antibodies against icam1  (Thermo Fisher)

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: Mediators of Inflammation

Article Title: CXCL1-Triggered Interaction of LFA1 and ICAM1 Control Glucose-Induced Leukocyte Recruitment during Inflammation In Vivo

doi: 10.1155/2012/739176

Figure Lengend Snippet: Leukocyte adhesion (number of adherent cells/mm 2 ) in mouse cremaster muscle venules and leukocyte transmigration (perivascular neutrophils/mm 2 surface area) in cremaster muscle whole mounts in response to glucose during trauma-induced inflammation. (a) Leukocyte adhesion was observed in trauma-stimulated cremaster muscle venules of wild-type mice (64 venules in 16 mice), LFA1-KO mice (19–25 venules in 4 mice), and ICAM1-KO mice (18–22 venules in 4 mice) before and during 15 minutes after intravenous injection of 0.5 g/kg glucose. To dissect the role of the chemokine pathway in this model, WT mice were either pretreated with 4 μ g PTx 3 h prior to cremaster exteriorization and glucose stimulation or coinjected with 600 ng CXCL1 (KC) and glucose and compared to injection of normal saline. WT mice injected with the equivalent volume of normal saline (18 venules in 4 mice) and the biologically inactive L-glucose (13–28 venules in 5 mice) served as controls. (b) Giemsa-stained cremaster muscle whole mounts of WT mice were analyzed for the number of perivascular neutrophils (per mm 2 surface area) after injection of glucose or normal saline in the trauma model). In parallel, leukocyte extravasation was quantified in glucose-treated LFA1-KO, ICAM1-KO mice, and WT mice pretreated with 4 μ g PTx 3 h prior to cremaster muscle exteriorization or coinjected with 600 ng CXCL1. To further illustrate the results representative micrographs of cremaster muscle whole mounts are shown in (c)–(i). Reference bar is shown in (c). Arrows point to extravasated leukocytes. All values are presented as mean ± SEM. Significances are indicated by the asterisks (* P < 0.05 versus WT control mice).

Article Snippet: Briefly, after insertion of a catheter into the carotid artery, primary antibodies against ICAM1 (YN-1, monoclonal rat anti-mouse; 30 μ g/mouse, ATCC, Wesel, Germany) were systemically injected and incubated for 10 minutes.

Techniques: Transmigration Assay, Injection, Saline, Staining, Control

Journal: Mediators of Inflammation

Article Title: CXCL1-Triggered Interaction of LFA1 and ICAM1 Control Glucose-Induced Leukocyte Recruitment during Inflammation In Vivo

doi: 10.1155/2012/739176

Figure Lengend Snippet: Hemodynamic and microvascular parameters of cremaster muscle venules in ICAM1-KO, LFA1-KO, and PTx-pretreated or KC-coinjected WT mice before and after injection 0,5 g/kg glucose in the trauma model. Vessel diameter, centerline velocity, and wall shear rate are displayed before and after the intravenous administration of glucose. Experiments are presented as mean ± SEM; n.s.: not significant stating no differences of hemodynamic parameters among the different groups and during the time course after glucose/saline injection.

Article Snippet: Briefly, after insertion of a catheter into the carotid artery, primary antibodies against ICAM1 (YN-1, monoclonal rat anti-mouse; 30 μ g/mouse, ATCC, Wesel, Germany) were systemically injected and incubated for 10 minutes.

Techniques: Injection, Shear

Journal: Mediators of Inflammation

Article Title: CXCL1-Triggered Interaction of LFA1 and ICAM1 Control Glucose-Induced Leukocyte Recruitment during Inflammation In Vivo

doi: 10.1155/2012/739176

Figure Lengend Snippet: Comparison of adherent cells in microflow chambers perfused with glucose or saline. Microflow chambers were coated with P selectin, ICAM1, and KC and constantly perfused with isolated glucose-stimulated or saline control bone marrow neutrophils of LysEGFP mice ( n = 4). Adhesion of leukocytes in fully coated flow chambers was compared to uncoated flow chambers.

Article Snippet: Briefly, after insertion of a catheter into the carotid artery, primary antibodies against ICAM1 (YN-1, monoclonal rat anti-mouse; 30 μ g/mouse, ATCC, Wesel, Germany) were systemically injected and incubated for 10 minutes.

Techniques: Comparison, Saline, Isolation, Control

Journal: Mediators of Inflammation

Article Title: CXCL1-Triggered Interaction of LFA1 and ICAM1 Control Glucose-Induced Leukocyte Recruitment during Inflammation In Vivo

doi: 10.1155/2012/739176

Figure Lengend Snippet: Glucose-induced expression of ICAM1 in surgically prepared cremaster muscle venules. Immunostaining was conducted to assess endothelial expression of ICAM1 in postcapillary venules of cremaster muscles obtained 15 minutes after exteriorization and injection of 0.5 g/kg glucose or the equivalent volume of normal saline. Application of primary antibody was performed i.v. before harvesting the cremaster muscle in order to stain ICAM1 on the endothelial surface. Biotinylated secondary antibody, peroxidase-conjugated streptavidin, and diaminobenzidine (DAB) were used to detect endothelial expression of ICAM1 as brown signal. Counter-staining was performed by Mayer's hemalaun. Intensity of venular anti-ICAM1 immunostaining during trauma-induced inflammation was analyzed semiquantitatively and presented as mean ± SEM (a; 0 = no, 1 = weak, 2 = medium, 3 = strong signal; at least 3 mice/group). In addition, representative images of saline- and glucose-treated mice are depicted in (b) and (c), respectively. Reference bar is shown in (b) and represents 30 μ m. Significant differences ( P < 0.05) are indicated by the asterisk.

Article Snippet: Briefly, after insertion of a catheter into the carotid artery, primary antibodies against ICAM1 (YN-1, monoclonal rat anti-mouse; 30 μ g/mouse, ATCC, Wesel, Germany) were systemically injected and incubated for 10 minutes.

Techniques: Expressing, Immunostaining, Muscles, Injection, Saline, Staining