Journal:

Article Title: Topoisomerase II? Negatively Modulates Retinoic Acid Receptor ? Function: a Novel Mechanism of Retinoic Acid Resistance ▿

doi: 10.1128/MCB.01576-07

Figure Lengend Snippet: TopoIIβ negatively regulates RA target genes. Real-time PCR analysis of RARβ, RIGI, HOXA1, and ICAM1 mRNA levels for the NB4 cell line (A) and the NB4-MR2 cell line (B), using the GAPDH gene as a reference gene. (A) mRNA levels for RARβ,

Article Snippet: The reaction was carried out at 42°C for 50 min in the presence of SuperScript II reverse transcriptase (Invitrogen). cDNA was amplified for RARβ and RIGI by real-time PCR analysis (ABI Prism7500; Applied Biosystems) using hybridization probes. cDNA was amplified for ICAM1 and HOXA1, by real-time PCR analysis (ABI Prism7500; Applied Biosystems) using primer sets as follows: for ICAM1, 5′ TGG CCC TCC ATA GAC ATG TGT 3′ (sense) and 5′ TGG CAT CCG TCA GGA AGT G 3′ (antisense); and for HOXA1, 5′ ACC CCG CCA GGA AAC G 3′ (sense) and 5′ GGC GAA GAG CTG GAC TTC TCT 3′ (antisense).

Techniques: Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages

doi: 10.1371/journal.pone.0206759

Figure Lengend Snippet: ICAM-1 and CD11c co-localize during exosome-macrophage interactions. (A) Immunoblotting analysis reveals exosomes from the pancreatic cancer cell lines AsPC-1 and BxPC-3 are enriched in the surface-exposed proteins ICAM-1 and EpCAM, while CD9 is more broadly expressed. The pan-exosomal marker flotillin-1 is used as a loading control. (B) Co-localization of exosome proteins and macrophage proteins is demonstrated by immunostaining for the exosome marker ICAM-1 (green, panel 1) or the macrophage marker CD11c (red, panel 2) after mixing AsPC-1 exosomes with THP-1-derived, non-polarized macrophages. Panel 3 is a merged image showing co-localization of ICAM-1 and CD11c staining (yellow, indicated by an arrow), and is suggestive of ICAM-1 and CD11c protein-protein interaction. THP-1-derived macrophages not mixed with exosomes (cells only) show little ICAM-1 staining (green, panel 4), suggesting exosomes were the main source of the ICAM-1 signal. In the absence of exosomes, more CD11c is associated with the THP-1 cell surface (red, panel 5), and merged images of THP-1 only staining show few areas of signal co-localization (panel 6, yellow). Scale bar = 3.0 μm (C) Quantitation of ICAM-1:CD11c co-localized signal in THP-1 cells mixed with AsPC-1 exosomes or in THP-1 cells alone. *** p

Article Snippet: When only THP-1-derived macrophages (no exosomes) were visualized, little ICAM-1 signal was observed, suggesting that the ICAM-1 was mainly associated with the AsPC-1 exosomes and not with the THP-1 macrophages, and the CD11c signal remained at the cell surface.

Techniques: Marker, Immunostaining, Derivative Assay, Staining, Quantitation Assay

Journal: Clinical and Developmental Immunology

Article Title: Expression and Function of the Homeostatic Molecule Del-1 in Endothelial Cells and the Periodontal Tissue

doi: 10.1155/2013/617809

Figure Lengend Snippet: Del-1 inhibits ICAM-1-dependent chemokine production by neutrophils. (a)–(c) ATRA-differentiated HL-60 neutrophils were cultured for 24 hours on plates coated with Fc protein control, ICAM-1-Fc, or Del-1-Fc, and induction of release of CCL3 (a), CXCL2 (b), and TNF (c) was assayed by ELISA. (d)-(e) ATRA-differentiated HL-60 neutrophils were cultured for 24 hours on ICAM-1-Fc-coated plates in the presence of the indicated concentrations of soluble Del-1, and induction of release of CCL3 (d) and CXCL2 (e) was assayed by ELISA. (f)-(g) ATRA-differentiated HL-60 neutrophils were stimulated for 24 hours with 1 μ g/mL Pam 3 CSK 4 or 0.1 μ g/mL LPS, in the absence or presence of the indicated concentrations of Del-1, and induction of release of CCL3 (f) and CXCL2 (g) was assayed by ELISA. Data are means ± SD ((a)–(e), n ≥ 4 and (f)-(g), n = 2 sets of HL-60 cells). * P

Article Snippet: Firm adhesion and crawling are primarily mediated by leukocyte integrins that interact with endothelial counterreceptors such as VCAM-1 and ICAM-1 [ , , ].

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Clinical and Developmental Immunology

Article Title: Expression and Function of the Homeostatic Molecule Del-1 in Endothelial Cells and the Periodontal Tissue

doi: 10.1155/2013/617809

Figure Lengend Snippet: Regulation of Del-1 versus ICAM-1 and VCAM-1 by inflammatory stimuli. HUVEC were cultured for the indicated time intervals with 10 ng/mL TNF (a), 5 μ g/mL serum amyloid A (SAA; (b)), P. gingivalis (Pg; MOI = 10 : 1) (c), Pam 3 CSK 4 or LPS (both at 0.5 μ g/mL; (d)), 10 ng/mL IFN γ (e), or IL-17A or IL-17F (both at 10 ng/mL; (f)) and assayed for Del-1, ICAM-1, and VCAM-1 mRNA expression. Results were normalized to those of GAPDH mRNA and expressed as fold change in transcript levels relative to those of medium-treated cells at 2 hours (HRS), the average value of which was taken as 1. The medium-treated groups in (b) and (c) are the same (SAA and Pg were tested together but were separated in the graphs for enhanced clarity). Data are means ± SD of duplicate determinations from one of three independent sets of experiments that yielded similar results.

Article Snippet: Firm adhesion and crawling are primarily mediated by leukocyte integrins that interact with endothelial counterreceptors such as VCAM-1 and ICAM-1 [ , , ].

Techniques: Cell Culture, Expressing

Journal: Clinical and Developmental Immunology

Article Title: Expression and Function of the Homeostatic Molecule Del-1 in Endothelial Cells and the Periodontal Tissue

doi: 10.1155/2013/617809

Figure Lengend Snippet: Expression of endothelial adhesion molecules in Del-1 deficiency. (a) Gingiva dissected from 16-week-old C57BL/6 Edil 3 +/+ (WT) and Edil 3 −/− mice were processed for quantitative real-time PCR analysis of mRNA expression of the indicated genes; results were normalized to those of GAPDH mRNA and expressed as fold change in Edil 3 −/− transcript levels relative to WT, the average value of which was taken as 1. (b) Sagittal sections of interdental gingiva from 16-week-old C57BL/6 WT or Edil 3 −/− mice were stained for ICAM-1, VCAM-1, or E-selectin and CD31, as indicated. Shown are representative overlays of differential interference contrast and fluorescent confocal images, with colocalization demonstrated in merged images (scale bar, 50 μ m). (c) The fluorescence intensities of the images shown here and of additional representative images from independent mice (5 per group) were quantified using ImageJ analysis. In (a) and (c), data are means ± SD ( n = 5 mice per group). No statistically significant differences were detected between Edil 3 −/− and WT littermate controls.

Article Snippet: Firm adhesion and crawling are primarily mediated by leukocyte integrins that interact with endothelial counterreceptors such as VCAM-1 and ICAM-1 [ , , ].

Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Staining, Fluorescence

Journal: Acta Neuropathologica Communications

Article Title: Hypoxic pre-conditioning suppresses experimental autoimmune encephalomyelitis by modifying multiple properties of blood vessels

doi: 10.1186/s40478-018-0590-5

Figure Lengend Snippet: CMH suppresses endothelial VCAM-1 and ICAM-1 expression during EAE. a and c . Frozen sections of lumbar spinal cord taken from disease-free, EAE-normoxia or EAE-CMH mice at the peak symptomatic phase of EAE were stained for CD31 (AlexaFluor-488) and VCAM-1 (Cy-3) in panel A or CD31 (Cy-3) and ICAM-1 (AlexaFluor-488) in panel C. Scale bar = 100 μm (inset, scale bar = 50 μm). b and d . Quantification of VCAM-1 ( b ) and ICAM-1 expression ( d ). Results are expressed as the mean ± SEM ( n = 6 mice/group). Note that CMH markedly suppressed vascular expression of VCAM-1 and ICAM-1. ** p

Article Snippet: Hamster monoclonal antibodies used included CD31 (clone 2H8) from Abcam (Cambridge, MA) and ICAM-1 (clone 3E2) from BD Pharmingen.

Techniques: Expressing, Mouse Assay, Staining

Journal: Journal of Ophthalmology

Article Title: Panretinal Photocoagulation Using Short-Pulse Laser Induces Less Inflammation and Macular Thickening in Patients with Diabetic Retinopathy

doi: 10.1155/2017/8530261

Figure Lengend Snippet: Alteration in the vitreous levels of inflammatory and angiogenic cytokines after panretinal photocoagulation (PRP) using the conventional laser with 500 spots (Conv 500s), short-pulse laser (SPL) with 500 spots (SPL 500), and SPL with 1000 spots (SPL 1000s). Concentrations of VEGF, IL-6, ICAM-1, and MCP-1 in the vitreous body were measured before and on 1, 7, and 14 days after PRP. Statistically significant difference as compared to the Conv group ( ∗ p

Article Snippet: Measurements of cytokine levels were carried out with a sandwich enzyme-linked immunosorbent assay using a commercially available kit (VEGF and IL-6; Cusabio Biotech Co. Ltd., Hubei, China, MCP-1; NEO Group Inc., MA, USA, ICAM-1; Cloud-Clone Corp., TX, USA) in accordance with the manufacturer's protocol.

Techniques:

Journal: PLoS ONE

Article Title: Effects of TLR Agonists on the Hypoxia-Regulated Transcription Factor HIF-1? and Dendritic Cell Maturation under Normoxic Conditions

doi: 10.1371/journal.pone.0010983

Figure Lengend Snippet: Effect of Digoxin (50 nM or 100 nM) on phenotypic maturation of MoDC. Monocytes isolated from buffy coat were cultured in the presence of GM-CSF and IL-4 for 6 days. MoDC were incubated with Digoxin for one hour, afterwards cells were stimulated for 24 hours with LPS (1 µg/ml), HA (20 µg/ml) and LTA (5 µg/ml). Cells were washed and analyzed for the expression of CD40 (A), CD80 (B), CD86 (C) and ICAM-1 (D). Y-axis shows the median fluorescence intensity (MFI). Mean values ± SD of 3 experiments with cells from different donors are shown.

Article Snippet: FACS analysis and cell viability Cells were incubated with FITC- or PE-labeled monoclonal antibody (mAb) against CD80, CD86 (BD, Franklin Lakes, NJ, USA), isotype control IgG1 (BD), or unlabeled mAb against CD40, ICAM-1 (Diaclone, Besançon, France) followed by a FITC-labeled polyclonal goat anti-mouse IgG (Sigma).

Techniques: Isolation, Cell Culture, Incubation, Expressing, Fluorescence

Journal: PLoS ONE

Article Title: Effects of TLR Agonists on the Hypoxia-Regulated Transcription Factor HIF-1? and Dendritic Cell Maturation under Normoxic Conditions

doi: 10.1371/journal.pone.0010983

Figure Lengend Snippet: Effect of YC-1 on phenotypic maturation of MoDC. Monocytes isolated from buffy coat were cultured in the presence of GM-CSF and IL-4 for 6 days. MoDC were incubated with YC-1 (100 µM) for 5 min, afterwards cells were stimulated for 24 hours with LPS (1 µg/ml), HA µ20 (g/ml) and LTA (5 µg/ml). Cells were washed and analyzed for the expression of CD40 (A), CD80 (B), CD86 (C) and ICAM-1 (D). Y-axis shows the median fluorescence intensity (MFI). Mean values ± SD of 4 experiments with cells from different donors are shown. *p

Article Snippet: FACS analysis and cell viability Cells were incubated with FITC- or PE-labeled monoclonal antibody (mAb) against CD80, CD86 (BD, Franklin Lakes, NJ, USA), isotype control IgG1 (BD), or unlabeled mAb against CD40, ICAM-1 (Diaclone, Besançon, France) followed by a FITC-labeled polyclonal goat anti-mouse IgG (Sigma).

Techniques: Isolation, Cell Culture, Incubation, Expressing, Fluorescence

Journal: PLoS ONE

Article Title: Effects of TLR Agonists on the Hypoxia-Regulated Transcription Factor HIF-1? and Dendritic Cell Maturation under Normoxic Conditions

doi: 10.1371/journal.pone.0010983

Figure Lengend Snippet: Effect of CTM on phenotypic maturation of MoDC. Monocytes isolated from buffy coat were cultured in the presence of GM-CSF and IL-4 for 6 days. Inhibition of HIF-1α expression by CTM inhibited maturation of MoDC in response to LPS. MoDC were incubated with CTM (200 nM) for 3 hours, afterwards cells were stimulated for 24 hours with LPS (1 µg/ml), HA (20 µg/ml) and LTA (5 µg/ml). Cells were washed and analyzed for the expression of CD40 (A), CD80 (B), CD86 (C) and ICAM-1 (D). Y-axis shows the median fluorescence intensity (MFI). Mean values ± SD of 6 experiments with cells from different donors are shown. *p

Article Snippet: FACS analysis and cell viability Cells were incubated with FITC- or PE-labeled monoclonal antibody (mAb) against CD80, CD86 (BD, Franklin Lakes, NJ, USA), isotype control IgG1 (BD), or unlabeled mAb against CD40, ICAM-1 (Diaclone, Besançon, France) followed by a FITC-labeled polyclonal goat anti-mouse IgG (Sigma).

Techniques: Isolation, Cell Culture, Inhibition, Expressing, Incubation, Fluorescence

Journal: Clinical and Experimental Immunology

Article Title: Activation of epithelial and myoepithelial cells in the salivary glands of patients with Sj?gren's syndrome: high expression of intercellular adhesion molecule-1 (ICAM.1) in biopsy specimens and cultured cells

doi: 10.1046/j.1365-2249.2001.01500.x

Figure Lengend Snippet: a, Representative example of constitutive ICAM.1 expression (3+ staining intensity score, SI) by a SGEC line established from a patient with primary SS and cultured in SEM medium (Original magnification × 46). b, Low spontaneous ICAM.1 expression (SI: 1+) by a SEM-cultured SGEC line obtained from a control patient (Original magnification × 46). c, Down-regulation of spontaneous ICAM.1 expression by the SGEC line shown in (a), upon cultivation in the serum-free, low calcium-containing KBM medium. Only a small population of flattened-cuboidal epithelial cells (arrowheads) retains strong levels of ICAM.1 expression (Original magnification × 46). d, Induction of strong expression of ICAM.1 by the KBM-cultured SGEC line shown in (c), following treatment with IFNα (Original magnification × 46). e, Representative example of ICAM.1 expression and morphological features attained by SGEC upon transfer of cells from KBM to calcium-supplemented KBM culture medium (CS-KBM). Cultivation in CS-KBM restores the spontaneous expression of ICAM.1 at a significant degree (SI: 2+) and leads to the acquisition of morphological characteristics largely identical to those of cells cultured in SEM (tightly connected, large and flattened cells) (Original magnification × 46). f, Weak basal expression of ICAM.1 (SI: 1+) by a control SGEC line cultured in CS-KBM. Addition of calcium in KBM does not up-regulate the spontaneous ICAM.1 expression (Original magnification × 46).

Article Snippet: The down-regulation of ICAM.1 on SGEC lines cultivated in KBM was associated with the acquisition of morphological features indicative of less differentiated epithelial cells (i.e. round, without cell-to-cell contact) ( ).

Techniques: Expressing, Staining, Cell Culture

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Doinseunggitang Ameliorates Endothelial Dysfunction in Diabetic Atherosclerosis

doi: 10.1155/2013/783576

Figure Lengend Snippet: Effect of DYSGT on ET-1, ICAM-1, and PPAR- γ protein expression in the aorta of ApoE KO mice. Western blots and corresponding densitometric analyses of ET-1, ICAM-1, and PPAR- γ in aortic tissue. Values are expressed as mean ± S.E. ( n = 4); ** P

Article Snippet: Similarly, in WTD-fed ApoE KO mice, immunofluorescence of the aorta tissues showed that ICAM-1 and ET-1 expression were decreased in rosiglitazone and DYSGT groups compared with ApoE KO group ( ).

Techniques: Expressing, Mouse Assay, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Doinseunggitang Ameliorates Endothelial Dysfunction in Diabetic Atherosclerosis

doi: 10.1155/2013/783576

Figure Lengend Snippet: Immunofluorescence staining of (a) ICAM-1 and (b) ET-1 in the aorta. Representative histological sections are thoracic aorta of Cont., ApoE KO mice, ApoE mice treated with rosiglitazone, and ApoE mice treated with DYSGT incubated with anti-ICAM-1, anti-ET-1 antibodies, respectively. Original magnification: 100x.

Article Snippet: Similarly, in WTD-fed ApoE KO mice, immunofluorescence of the aorta tissues showed that ICAM-1 and ET-1 expression were decreased in rosiglitazone and DYSGT groups compared with ApoE KO group ( ).

Techniques: Immunofluorescence, Staining, Mouse Assay, Incubation

Journal: Journal of Leukocyte Biology

Article Title: WASp-dependent actin cytoskeleton stability at the dendritic cell immunological synapse is required for extensive, functional T cell contacts

doi: 10.1189/jlb.2A0215-050RR

Figure Lengend Snippet: The WASp-dependent DC actin cytoskeleton contributes to correct organization of adhesion molecules and formation of an extensive cell:cell contact. (A) WT, WASKO, and Y293F DCs (yellow/orange), pulsed with OVA, were cocultured with OT-II T cells (blue/green). After 1 h, conjugates were fixed, processed, and imaged using Gatan 3View. Isosurface reconstructions were created in Amira. (Lower) Conjugates with T cell removed to visualize the contact interface. (B) Quantification of DC:T cell contact surface area as a percentage of T cell surface. A minimum of 10 conjugates was analyzed per group. Unpaired t test was used to test significance among DC types; *** P (WT + WASKO) = 0.0005, P (WASKO + Y293F) = 0.0002; ns P (WT + Y293F) = 0.3153. (C and D) DCs expressing ICAM-1-GFP (green) were cocultured with T cells (red; anti-TCR immunostain) for 45 min and fixed. Images represent a slice cutting through the synapse. Polarization ratios of ICAM-1 on the DC side were calculated by measuring fluorescence intensity at the synapse normalized to whole cell. Original scale bars, 5 μm. Unpaired t test was used to test significance among DC types; ** P (WT + WASKO) = 0.0026; ns, P (WT + Y293F) = 0.1875, P (WASKO + Y293F) = 0.0610. DIC, Differential interference contrast. (E) Total surface ICAM-1 was measured by flow cytometry in immature and LPS-matured BMDCs, gated on CD11c-positive cells. Means and sd are shown from 3 independent experiments. MFI, Mean fluorescence intensity. (F) Total polymerized actin was measured using phalloidin in permeabilized, immature and LPS-matured BMDCs. Staining was performed in mixed population samples using CFSE labeling. Bars represent means and sd from 3 experiments. *** P (WT + WASKO) = 0.0010, * P (WT + Y293F) = 0.0472, * P (WASKO + Y293F) = 0.0407.

Article Snippet: With the use of supported planar lipid bilayers incorporating anti-ICAM-1 and anti-MHC class II antibodies, the dendritic cell actin cytoskeleton organized into recognizable synaptic structures but interestingly, formed Wiskott-Aldrich syndrome protein-dependent podosomes within this area.

Techniques: Expressing, Fluorescence, Flow Cytometry, Cytometry, Staining, Labeling

Journal: Journal of Leukocyte Biology

Article Title: WASp-dependent actin cytoskeleton stability at the dendritic cell immunological synapse is required for extensive, functional T cell contacts

doi: 10.1189/jlb.2A0215-050RR

Figure Lengend Snippet: The development of a novel imaging system of the DC synapse. (A) Three different lipid bilayer compositions were designed to mimic a DC:T cell synapse. A biotinylated α-MHC II-Cy5-conjugated antibody was incorporated to replace TCR interaction ( Fig. 2 ). Alternatively, both α-MHC II-Cy5 and α-ICAM-1 antibodies were added to replicate adhesion forces (A and E). α-ICAM-1 alone was used in D. (B) ICAM-1-GFP-expressing DCs interacting with an α-MHC II bilayer were fixed after 20 min and imaged. Original scale bars, 5 μm. Fluorescence intensity of ICAM-1-GFP and MHC II-Cy5 is plotted along the cell diameter, showing differential distribution in WT and WASKO DC. (C) Number of cells exhibiting radial symmetry, as a percentage of cells interacting with the α-MHC II bilayer. A minimum of 30 cells per strain was analyzed. Cells with radial symmetry were defined as having at least 3 different diameter cross-sections showing plots similar to WT DC in B. * P = 0.0232; ** P = 0.0092; ns, P = 0.0572. (D) WT, WASKO, and Y293F DCs interacting with the α-MHC II-Cy5 (red) bilayer were fixed and stained with phalloidin (blue). Original scale bars, 5 μm. (E) Three parameters were measured by use of ImageJ “Measure” and “Analyze particles” functions in cells interacting with an α-MHC II-Cy5 bilayer: average actin intensity across the contact; MHC II area as a percentage of the total contact area (actin); and number of peripheral microclusters (MC) per cell (size

Article Snippet: With the use of supported planar lipid bilayers incorporating anti-ICAM-1 and anti-MHC class II antibodies, the dendritic cell actin cytoskeleton organized into recognizable synaptic structures but interestingly, formed Wiskott-Aldrich syndrome protein-dependent podosomes within this area.

Techniques: Imaging, Expressing, Fluorescence, Staining

Journal: Journal of Leukocyte Biology

Article Title: WASp-dependent actin cytoskeleton stability at the dendritic cell immunological synapse is required for extensive, functional T cell contacts

doi: 10.1189/jlb.2A0215-050RR

Figure Lengend Snippet: Novel actin organization at the DC synapse. (A) WT, WASKO, and Y293F DCs interacting with an α-MHC II-Cy5 (red) and α-ICAM-1 bilayer were fixed and stained with phalloidin (blue). Original scale bars, 5 μm. (B) The polymerized actin was stained with phalloidin, and fluorescent intensity at the contact site as well as total actin area was quantified at the 4 time points. A minimum of 100 cells was analyzed in 4 experiments; means and sem are plotted. For actin intensity: *5 min: P (WT + WASKO) = 0.0113, P (WT + Y293F) = 0.0428; ***15 min: P (WT + WASKO) = 0.0005, P (WASKO + Y293F) = 0.0256; ***30 min: P (WT + WASKO) = 0.0007; ***60 min: P (WT + WASKO) = 0.0009. For total actin area: *15 min: P (WT + WASKO) = 0.0346. (C) Percentage of WT, WASKO, and Y293F DCs forming podosomes on an α-MHC II and α-ICAM-1 bilayer. A minimum of 400 cells was analyzed at each time point. “Actin clusters” are irregular, high-intensity actin structures, similar to those in WASKO cells at 30 and 60 min (A). (D) WT DC contacting an α-ICAM-1-only bilayer. Position of cells is depicted by DAPI staining (white). Phalloidin staining (blue) shows podosome rosettes. (E) The proportion of WT cells forming rings of podosomes in the 3 bilayer conditions is quantified. Means and sem from 3 experiments are shown; a minimum of 300 cells was analyzed. * P = 0.0241, ** P = 0.0073 (F) WT DC contacting an α-MHC II-Cy5 and α-ICAM-1 bilayer, showing actin-rich podosomes (blue) and immunofluorescent staining (yellow): capping protein (F-actin capping protein, α subunit; upper), vinculin (lower). Colocalization of F-actin capping protein and actin produces a white overlay; 36 WT DCs were analyzed to calculate colocalization. Pearson correlation coefficient = 0.442 ± 0.14; Mander’s overlap coefficient = 0.777 ± 0.04. Original scale bars, 5 μm. A 3× zoom is shown to the right. (G) DCs were seeded on 2 different bilayers and on fibronectin (50 μg/ml) and fixed at set intervals. Diameter of the podosome actin cores was measured in ImageJ; > 100 podosomes were measured for each condition. Synapse podosomes did not change significantly over time and showed a similar size to those formed on the ventral side of cells adhering to fibronectin.

Article Snippet: With the use of supported planar lipid bilayers incorporating anti-ICAM-1 and anti-MHC class II antibodies, the dendritic cell actin cytoskeleton organized into recognizable synaptic structures but interestingly, formed Wiskott-Aldrich syndrome protein-dependent podosomes within this area.

Techniques: Staining

Journal: Infection and Immunity

Article Title: Porphyromonas gingivalis Fimbriae Use ?2 Integrin (CD11/CD18) on Mouse Peritoneal Macrophages as a Cellular Receptor, and the CD18 ? Chain Plays a Functional Role in Fimbrial Signaling

doi:

Figure Lengend Snippet: ICAM-1 inhibits the fimbria-induced expression of IL-1β and TNF-α genes in peritoneal macrophages. (A) Peritoneal macrophages in each well of a microculture plate were pretreated or not pretreated for 3 h with ICAM-1 at 50 U/ml and then incubated for 1 h with or without fimbriae (4 μg of protein/ml). After incubation, total RNAs were prepared, and Northern blot analysis was performed with IL-1β, TNF-α, and β-actin cDNAs as probes. (B) Quantification of IL-1β and TNF-α gene expression in panel A was done by densitometry, and the results are expressed as percent inhibition. An identical experiment performed independently gave similar results.

Article Snippet: Therefore, the cells were pretreated or not pretreated with ICAM-1 at 50 U/ml for 3 h before the addition of fimbriae and then were incubated for 1 h in the absence or presence of fimbriae.

Techniques: Expressing, Incubation, Northern Blot, Inhibition

Journal: Infection and Immunity

Article Title: Porphyromonas gingivalis Fimbriae Use ?2 Integrin (CD11/CD18) on Mouse Peritoneal Macrophages as a Cellular Receptor, and the CD18 ? Chain Plays a Functional Role in Fimbrial Signaling

doi:

Figure Lengend Snippet: ICAM-1 inhibits the binding of 125 I-labeled fimbriae to mouse peritoneal macrophages. 125 I-labeled fimbriae (1 μg of protein) were treated or not treated with ICAM-1 (A) or laminin (B) at the indicated doses in each well of a microculture plate containing formalin-fixed peritoneal macrophages. The binding of 125 I-labeled fimbriae to cells was measured after 4 h. Each assay was carried out in triplicate. The results are expressed as the mean ± SD percent inhibition of binding of 125 I-labeled fimbriae (40,050 ± 2,171 cpm) in the absence of test samples. An identical experiment performed independently gave similar results.

Article Snippet: Therefore, the cells were pretreated or not pretreated with ICAM-1 at 50 U/ml for 3 h before the addition of fimbriae and then were incubated for 1 h in the absence or presence of fimbriae.

Techniques: Binding Assay, Labeling, Inhibition

Journal: Oncotarget

Article Title: Restoration of immune surface molecules in Kaposi sarcoma-associated herpes virus infected cells by lenalidomide and pomalidomide

doi: 10.18632/oncotarget.17960

Figure Lengend Snippet: Effect of Pom on MHC-I (HLA-A, B and C), ICAM and B7-2 mRNA gene expression in BCBL-1 cells induced with butyrate BCBL-1 cells were treated with control (DMSO) or Pom (1 μM) for 24 h followed by treatment with PBS or butyrate for an additional 24 h. Total mRNA was isolated and analyzed for expression levels by real time QT-PCR and was normalized to 18S levels. Shown are the fold changes in mRNA expression for total A . HLA and HLA A, B and C alleles following treatment and B . ICAM-1 and B7-2. Values are the average +/- standard deviation of four independent experiments. *** P

Article Snippet: Primers for ICAM-1 and B7-2 were from Biorad (ICAM-1: 10025636, qHsaCED0004281) and (B7-2: 10025636, qHsaCED0043530 (Hercules, CA).

Techniques: Expressing, Isolation, Polymerase Chain Reaction, Standard Deviation

Journal: Oncotarget

Article Title: Restoration of immune surface molecules in Kaposi sarcoma-associated herpes virus infected cells by lenalidomide and pomalidomide

doi: 10.18632/oncotarget.17960

Figure Lengend Snippet: Restoration of ICAM-1 and B7-2 but not ICAM-3 expression in BCBL-1 cells treated with Pom A . ICAM-1 expression in BCBL-1 cells and in B . BJAB cells as determined by FACS after pretreatment for 48 h with DMSO vehicle control or Pom (1 μM). C . B7-2 expression in BCBL-1 cells and in D . BJAB cells as determined by FACS after pretreatment for 48 h with DMSO vehicle control or Pom (1 μM). E . ICAM-3 expression in BCBL-1 cells and in F . BJAB cells as determined by FACS after pretreatment for 48 h with DMSO vehicle control or Pom (1 μM). The data shows a representative experiment of four independent experiments for A . and C . and two independent experiments for B ., D ., and F . with similar results. The grey shading shows the isotype control.

Article Snippet: Primers for ICAM-1 and B7-2 were from Biorad (ICAM-1: 10025636, qHsaCED0004281) and (B7-2: 10025636, qHsaCED0043530 (Hercules, CA).

Techniques: Expressing, FACS

Journal: Oncotarget

Article Title: Restoration of immune surface molecules in Kaposi sarcoma-associated herpes virus infected cells by lenalidomide and pomalidomide

doi: 10.18632/oncotarget.17960

Figure Lengend Snippet: Effect of Len on ICAM-1 and B7-2 expression in BCBL-1 cells, effect of Pom on MHC-II expression in BCBL-1 and BJAB cells, and effect of Pom on ICAM-1 and B7-2 in JSC-1 cells A . ICAM-1 expression and B . B7-2 expression in BCBL-1 cells pretreated for 48 h with DMSO vehicle control or Pom (1 μM) as determined by FACS. C . ICAM-1 expression D . B7-2 expression in JSC-1 cells pretreated for 48 h with DMSO vehicle control or Pom (0.2 μM) as determined by FACS. E . MHC-II expression in BCBL-1 cells or F . BJAB cells pretreated for 48 h with DMSO vehicle control or Pom (1 μM) as determined by FACS. The data is representative of two independent experiments in A, one experiment in B-D, and two experiments in E and F. The grey shading shows the isotype control.

Article Snippet: Primers for ICAM-1 and B7-2 were from Biorad (ICAM-1: 10025636, qHsaCED0004281) and (B7-2: 10025636, qHsaCED0043530 (Hercules, CA).

Techniques: Expressing, FACS

Journal: Scientific Reports

Article Title: 9c11tCLA modulates 11t18:1 and 9t18:1 induced inflammations differently in human umbilical vein endothelial cells

doi: 10.1038/s41598-018-19729-9

Figure Lengend Snippet: Effect of 9t18:1 and 11t18:1 on gene expression of ICAM-1, VCAM-1 and IL-6 of leptin/9c11t-CLA treated HUVECs. ( A ) The effect of 11t18:1 on expression of ICAM-1, VCAM-1 and IL-6 in leptin treated HUVECs. HUVECs were treated or without leptin (75 nmol/L) for 24 h and then cultured with 11t18:1(25, 50, 100 μmol/L) for 24 h. ( B ) The effect of 9t18:1 on expression of ICAM-1, VCAM-1 and IL-6 in 9c11tCLA treated HUVECs. HUVECs were treated or non-treated with 9c11tCLA (5, 25, 50 μmol/L) and then cultured with 9t18:1 (100 μmol/L) for 24 h. ( C ) The effect of 11t18:1 + leptin on expression of ICAM-1, VCAM-1 and IL-6 in 9c11tCLA treated HUVECs. HUVECs were treated or non-treated with 9c11tCLA (5, 25, 50 μmol/L) and then cultured with the group of 11t18:1 (100 μmol/L) + leptin (75 nmol/L) for 24 h. ( D ) The effect of 9t18:1 on expression of ICAM-1, VCAM-1 and IL-6 of leptin treated HUVECs. HUVECs were treated or non-treated with leptin (75 nmol/L) and then cultured with 9t18:1 (100 μmol/L) and 11t18:1 (100 μmol/L) for 24 h. a–g Data were presented as mean ± SD, values not sharing a common superscript denote significant difference ( P

Article Snippet: Anti-ICAM-1, anti-p38, anti-p-p38, anti-SAPK/JNK, anti-p-SAPK/JNK, anti-ERK 1/2, anti-p-ERK1/2, anti-TLR4 and anti-SCD-1 were purchased from Cell Signaling Technology (USA).

Techniques: Expressing, Cell Culture

Journal: Scientific Reports

Article Title: 9c11tCLA modulates 11t18:1 and 9t18:1 induced inflammations differently in human umbilical vein endothelial cells

doi: 10.1038/s41598-018-19729-9

Figure Lengend Snippet: Effect of TAK242 on MAPKs phosphorylation in HUVECs treated with 9t18:1 and 11t18:1. ( A ) Effect of TAK242 on ICAM-1 expression. HUVECs were treated with TAK242 (0.5, 1, 1.5 μmol/L) for 30 min and then cultured with 9t18:1 for 24 h. ( B ) Effect of TAK242 on MAPKs phosphorylation in HUVECs treated with 9t18:1 and 11t18:1. HUVECs were treated with TAK242 (1 μmol/L) for 30 min and then cultured with TFA for 24 h. Values labeled with different letters in each set indicate significant differences ( p

Article Snippet: Anti-ICAM-1, anti-p38, anti-p-p38, anti-SAPK/JNK, anti-p-SAPK/JNK, anti-ERK 1/2, anti-p-ERK1/2, anti-TLR4 and anti-SCD-1 were purchased from Cell Signaling Technology (USA).

Techniques: Expressing, Cell Culture, Labeling

Journal: Journal of Histochemistry and Cytochemistry

Article Title: LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 Expression in Human Lymphatic Endothelium

doi: 10.1369/jhc.7A7299.2007

Figure Lengend Snippet: Induction of interleukin (IL)-6, IL-8, VCAM-1, and ICAM-1 mRNAs by LPS. ( A ) RT-PCR analysis. Expression of IL-6, IL-8, VCAM-1, and ICAM-1 mRNAs in LEC increased with LPS treatment for 24 hr. RT-PCR products for IL-6 mRNA were not detected in LEC but were detected in the cells with 100 ng/ml LPS, and were drastically increased in the cells with 1 μg/ml LPS. RT-PCR products for IL-8 mRNA were detected in LEC and increased in a dose-dependent manner. RT-PCR products for VCAM-1 mRNA were not detected in LEC but were detected in the cells with 10 ng/ml LPS, and drastically increased in the cells with 100 ng/ml LPS. RT-PCR products for ICAM-1 mRNA were detected in LEC and increased in a dose-dependent manner. MW, molecular weight marker. ( B ) Real-time quantitative PCR analysis. LEC with 100 ng/ml and 10 μg/ml LPS showed 8.3- and 223.3-fold increases of IL-6 mRNA, 6.9- and 26.6-fold increases of IL-8 mRNA, 10.7- and 91.3-fold increases of VCAM-1 mRNA, and 84.5- and 390-fold increases of ICAM-1 mRNA, when compared with untreated cells. *Not significantly different from the untreated cells.

Article Snippet: These findings suggest that human lymphatic endothelium has the ability to enhance the expression of VCAM-1 and ICAM-1 in response to LPS stimulation.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Molecular Weight, Marker, Real-time Polymerase Chain Reaction

Journal: Journal of Histochemistry and Cytochemistry

Article Title: LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 Expression in Human Lymphatic Endothelium

doi: 10.1369/jhc.7A7299.2007

Figure Lengend Snippet: Immunohistochemical analysis for the effect of LPS on vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) expression. Cells were visualized with rhodamine-conjugated Con A (Con A, in red). Reaction products with antibodies for VCAM-1 and ICAM-1 were visualized with an AF488-conjugated second antibody (AF488, in green) in human neonatal dermal LEC. Merged images indicate that expression of VCAM-1 was not observed in LEC but was observed in the cells with LPS, and that expression of ICAM-1 was observed in LEC and strongly in the cells with LPS. Bar = 100 μm.

Article Snippet: These findings suggest that human lymphatic endothelium has the ability to enhance the expression of VCAM-1 and ICAM-1 in response to LPS stimulation.

Techniques: Immunohistochemistry, Expressing

Journal: Journal of Histochemistry and Cytochemistry

Article Title: LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 Expression in Human Lymphatic Endothelium

doi: 10.1369/jhc.7A7299.2007

Figure Lengend Snippet: Effects of anti-TLR4, nobiletin, and caffeic acid phenethyl ester (CAPE) on LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 production. ( A ) RT-PCR analysis of LEC. RT-PCR products for LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 mRNA decreased in LEC pretreated with anti-TLR4, nobiletin, and CAPE when compared with LEC with only LPS. MW, molecular weight marker. ( B ) Real-time quantitative PCR analysis. The pretreatment of LEC by anti-TLR4, nobiletin, and CAPE suppressed LPS-induced mRNA production: IL-6 to 21.5%, 5.5%, and 5.2% levels; IL-8 to 25.4%, 28.9%, and 19.1% levels; VCAM-1 to 24.4%, 11.2%, and 20% levels; and ICAM-1 to 54.4%, 49.8%, and 42.3% levels when compared with LPS-stimulated cells not pretreated. ( C ) Enzyme-linked immunosorbent assay. The pretreatment of LEC by anti-TLR4, nobiletin, and CAPE suppressed LPS-induced protein production: IL-6 to 52.2%, 20%, and 26.7%; IL-8 to 69.3%, 67.9%, and 52.6%; VCAM-1 to 36.1%, 18.1%, and 30.1%; and ICAM-1 to 54.7%, 53.8%, and 45.3% when compared with LPS-stimulated cells not pretreated. Error bars indicate mean ± SEM. *Not significantly different.

Article Snippet: These findings suggest that human lymphatic endothelium has the ability to enhance the expression of VCAM-1 and ICAM-1 in response to LPS stimulation.

Techniques: Reverse Transcription Polymerase Chain Reaction, Molecular Weight, Marker, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Journal of Histochemistry and Cytochemistry

Article Title: LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 Expression in Human Lymphatic Endothelium

doi: 10.1369/jhc.7A7299.2007

Figure Lengend Snippet: Effect of TLR4-specific small interfering RNA (siRNA) introduction on LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 mRNA production. ( A ) RT-PCR analysis of LEC. RT-PCR products for IL-6, IL-8, VCAM-1, and ICAM-1 mRNA decreased in siRNA-transfected LEC with LPS when compared with LEC with only LPS, and with mock and LPS. MW, molecular weight marker. ( B ) Real-time quantitative PCR analysis. LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 mRNA amounts in siRNA-transfected LEC were ∼50% of LEC with only LPS, and of LEC with mock and LPS. Error bars indicate mean ± SEM. *Not significantly different.

Article Snippet: These findings suggest that human lymphatic endothelium has the ability to enhance the expression of VCAM-1 and ICAM-1 in response to LPS stimulation.

Techniques: Small Interfering RNA, Reverse Transcription Polymerase Chain Reaction, Transfection, Molecular Weight, Marker, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Characterization of Renal Injury and Inflammation in an Experimental Model of Intravascular Hemolysis

doi: 10.3389/fimmu.2018.00179

Figure Lengend Snippet: Hemolysis inducing renal injury. Kinetics of mRNA levels in renal tissue of (A) NGAL, (B) Kim-1, (C) Ki-67, (D) IL-6, (E) E-selectin, (F) P-selectin and (G) ICAM-1 at 1, 2, and 4 days after PBS or phenylhydrazine (PHZ) injection. (H) Human umbilical vein endothelial cells (HUVECs) were incubated for 24 h with increased concentration of PHZ in M199 medium, containing 20% fetal calf serum, and cell death was measured by double staining with annexin-V and DAPI by flow cytometry. (I–K) HUVECs were incubated for 24 h with increased concentration of PHZ or TNF-α as a positive control, in M199 medium, containing 20% fetal calf serum. E-selectin (I) , VCAM-1 (J) , and ICAM-1 (K) were measured by flow cytometry. Data are presented for 0.312-mg/mL PHZ, gating on live (annexin-V − , DAPI − cells, about 80% of the total cell population). Mean ± SEM, * p

Article Snippet: For the ICAM-1 and P-selectin staining, a goat anti-mouse PE antibody 1:100 (Beckman, IM0855)was used for 30 min at 4°C.

Techniques: Injection, Incubation, Concentration Assay, Double Staining, Flow Cytometry, Cytometry, Positive Control

Journal: Frontiers in Immunology

Article Title: Characterization of Renal Injury and Inflammation in an Experimental Model of Intravascular Hemolysis

doi: 10.3389/fimmu.2018.00179

Figure Lengend Snippet: The renal injury which is largely heme-independent. mRNA levels of NGAL (A) , Kim-1 (B) , and IL-6 (C) in renal tissue after injection of heme at day 2. Influence of hemopexin (Hx) on mRNA level in the phenylhydrazine (PHZ) model, (D) NGAL, (E) Kim-1, (F) IL-6, (G) E-selectin, (H) P-selectin, and (I) ICAM-1 at day 1 or day 4, as mentioned under the panels. * p

Article Snippet: For the ICAM-1 and P-selectin staining, a goat anti-mouse PE antibody 1:100 (Beckman, IM0855)was used for 30 min at 4°C.

Techniques: Injection

Journal: PLoS ONE

Article Title: Ex Vivo Evidence for the Contribution of Hemodynamic Shear Stress Abnormalities to the Early Pathogenesis of Calcific Bicuspid Aortic Valve Disease

doi: 10.1371/journal.pone.0048843

Figure Lengend Snippet: Effects of TAV and BAV WSS on endothelial activation. ICAM-1 and VCAM-1 immunostaining in porcine aortic valve leaflets subjected to TAV and BAV WSS (F: fibrosa; V: ventricularis; green: positively stained cells; blue: cell nuclei).

Article Snippet: Following the blocking step, the slides were then incubated overnight at 4°C in primary antibody in 2–10% blocker at the following dilutions: VCAM-1 (1∶50, Santa Cruz Biotechnology, Santa Cruz, CA), ICAM-1 (1∶50, Southern Biotech, Birmingham, AL), TGF-β1 (1∶25, Santa Cruz), BMP-4 (1∶150, Abcam, Cambridge, MA), cathepsin L (1∶25, Santa Cruz), cathepsin S (1∶25, Santa Cruz), MMP-2 (1∶100, EMD Millipore, Billerica, MA) MMP-9 (1∶100, EMD Millipore), TIMP-2 (1∶50, Santa Cruz) and osteocalcin (1∶150, EMD Millipore).

Techniques: Activation Assay, Immunostaining, Staining

Journal: Transplantation

Article Title: Porcine CMV Infection Is Associated with Early Rejection of Kidney Grafts in a Pig to Baboon Xenotransplantation Model

doi: 10.1097/TP.0000000000000232

Figure Lengend Snippet: Immunofluorescence staining with ICAM-1 (A) and anti-swine class II antibodies A) Comparison of ICAM1 expression in pCMV positive kidneys with negative kidneys following xenotransplantation. a–c were from pCMV negative donors and e-f were from pCMV positive donors: a and d) native swine kidneys; b and e) non-rejected kidney xenografts; c and f): rejected kidney xenografts. ICAM-1 expression (white arrows) is seen on the peritubular capillaries in pCMV positive kidneys (e and f) while pCMV negative rejected kidney had only a minimal expression of ICAM-1 (c). B) The comparison of class II DR expression in pCMV positive kidneys with negative kidneys following xenotransplantation. a–c were from pCMV negative donors and e-f were from pCMV positive donors: a and d): naive kidneys; b and e): non-rejected kidney grafts, c and f): rejected kidney grafts. Up-regulation of class II on peritubular capillaries and glomeruli (white arrows) in a rejected pCMV negative kidneys (c) as well as pCMV positive rejected kidneys (e and f).

Article Snippet: Immunohistochemistry was also performed on frozen sections using polyclonal antibodies reactive to ICAM-1, porcine MHC Class II, and IgG (polyclonal rabbit anti-human IgG (Dako, Denmark).

Techniques: Immunofluorescence, Staining, Expressing

Journal: Journal of Proteome Research

Article Title: Stable Isotope Dilution Multidimensional Liquid Chromatography-Tandem Mass Spectrometry for Pancreatic Cancer Serum Biomarker Discovery

doi: 10.1021/pr800904z

Figure Lengend Snippet: ICAM-1 concentrations in the six cancer and six control serum samples studied. Mean ICAM-1 concentration in cancer sera was significantly greater than in controls.

Article Snippet: Both ICAM-1 and BCAM concentrations were significantly elevated in the individual cancer serum samples compared with control samples (Figure and Figure , respectively).

Techniques: Concentration Assay

Journal: Journal of Proteome Research

Article Title: Stable Isotope Dilution Multidimensional Liquid Chromatography-Tandem Mass Spectrometry for Pancreatic Cancer Serum Biomarker Discovery

doi: 10.1021/pr800904z

Figure Lengend Snippet: ICAM-1 concentrations in ten cancer and ten control serum samples from an independent cohort. Mean ICAM-1 concentration in cancer sera was significantly greater than in controls.

Article Snippet: Both ICAM-1 and BCAM concentrations were significantly elevated in the individual cancer serum samples compared with control samples (Figure and Figure , respectively).

Techniques: Concentration Assay