Structured Review

GE Healthcare icam 1
Effect of inhibitors of signal transduction pathway on the protein expression levels of adhesion molecules in HUVEC stimulated with LPS. Endothelial cell was treated with 50µmol/l of JSH (NF-κB inhibitor), SB202190 (p38 MAPK inhibitor), PD98059 (MEK/ERK inhibitor), wortmannin (PI3-K inhibitor), and SP600125 (JNK inhibitor) for 1hr prior to LPS (1µg/ml) stimulation for 8 hr. Cell extracts were resolved on 8% SDS-polyacrylamide gel and Western blot analysis with the respective primary antibody against VCAM-1 and <t>ICAM-1.</t> β-tubulin was used as an internal control. The bar graph represents the amount of VCAM-1 and ICAM-1 estimated by image scanning and is expressed in arbitrary units. Values are means±SEM of 3 independent experiments. Statistical significance assessed by one-way ANOVA followed by Scheffe post-hoc test for multiple comparisons ( * p
Icam 1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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icam 1 - by Bioz Stars, 2020-09
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Images

1) Product Images from "Ginsenoside Rg2 Inhibits Lipopolysaccharide-Induced Adhesion Molecule Expression in Human Umbilical Vein Endothelial Cell"

Article Title: Ginsenoside Rg2 Inhibits Lipopolysaccharide-Induced Adhesion Molecule Expression in Human Umbilical Vein Endothelial Cell

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

doi: 10.4196/kjpp.2013.17.2.133

Effect of inhibitors of signal transduction pathway on the protein expression levels of adhesion molecules in HUVEC stimulated with LPS. Endothelial cell was treated with 50µmol/l of JSH (NF-κB inhibitor), SB202190 (p38 MAPK inhibitor), PD98059 (MEK/ERK inhibitor), wortmannin (PI3-K inhibitor), and SP600125 (JNK inhibitor) for 1hr prior to LPS (1µg/ml) stimulation for 8 hr. Cell extracts were resolved on 8% SDS-polyacrylamide gel and Western blot analysis with the respective primary antibody against VCAM-1 and ICAM-1. β-tubulin was used as an internal control. The bar graph represents the amount of VCAM-1 and ICAM-1 estimated by image scanning and is expressed in arbitrary units. Values are means±SEM of 3 independent experiments. Statistical significance assessed by one-way ANOVA followed by Scheffe post-hoc test for multiple comparisons ( * p
Figure Legend Snippet: Effect of inhibitors of signal transduction pathway on the protein expression levels of adhesion molecules in HUVEC stimulated with LPS. Endothelial cell was treated with 50µmol/l of JSH (NF-κB inhibitor), SB202190 (p38 MAPK inhibitor), PD98059 (MEK/ERK inhibitor), wortmannin (PI3-K inhibitor), and SP600125 (JNK inhibitor) for 1hr prior to LPS (1µg/ml) stimulation for 8 hr. Cell extracts were resolved on 8% SDS-polyacrylamide gel and Western blot analysis with the respective primary antibody against VCAM-1 and ICAM-1. β-tubulin was used as an internal control. The bar graph represents the amount of VCAM-1 and ICAM-1 estimated by image scanning and is expressed in arbitrary units. Values are means±SEM of 3 independent experiments. Statistical significance assessed by one-way ANOVA followed by Scheffe post-hoc test for multiple comparisons ( * p

Techniques Used: Transduction, Expressing, Western Blot

Effect of Rg2 on the protein expression levels of adhesion molecules in HUVEC stimulated with LPS. Endothelial cell was treated with 1, 10, 20, 50 and 100µmol/l of Rg2 for 1 hr prior to LPS (1µg/ml) stimulation for 8 hr. Cell extracts were resolved on 8% SDS-polyacrylamide gel and Western blot analysis with the respective primary antibody against VCAM-1 and ICAM-1. β-tubulin was used as an internal control. The bar graph represents the amount of VCAM-1 and ICAM-1 estimated by image scanning and is expressed in arbitrary units. Values are means±SEM of 3 independent experiments. Statistical significance assessed by one-way ANOVA followed by Scheffe post-hoc test for multiple comparisons ( ** p
Figure Legend Snippet: Effect of Rg2 on the protein expression levels of adhesion molecules in HUVEC stimulated with LPS. Endothelial cell was treated with 1, 10, 20, 50 and 100µmol/l of Rg2 for 1 hr prior to LPS (1µg/ml) stimulation for 8 hr. Cell extracts were resolved on 8% SDS-polyacrylamide gel and Western blot analysis with the respective primary antibody against VCAM-1 and ICAM-1. β-tubulin was used as an internal control. The bar graph represents the amount of VCAM-1 and ICAM-1 estimated by image scanning and is expressed in arbitrary units. Values are means±SEM of 3 independent experiments. Statistical significance assessed by one-way ANOVA followed by Scheffe post-hoc test for multiple comparisons ( ** p

Techniques Used: Expressing, Western Blot

2) Product Images from "Host cytosolic RNA sensing pathway promotes T Lymphocyte-mediated mycobacterial killing in macrophages"

Article Title: Host cytosolic RNA sensing pathway promotes T Lymphocyte-mediated mycobacterial killing in macrophages

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1008569

M . avium induces ICAM-1 production in macrophages through a RIG-I/MAVS/TBK1-dependent pathway. (A) Cytokine array analyzing indicated serum proteins in WT and Mavs -/- mice 3 weeks post M . avium intratracheal infection using a dose of 1x10 7 CFU/mouse. (B) Densitometry of serum proteins highlighted in (A). (C) qRT-PCR analysis of designated transcripts in uninfected or M . avium 104 infected WT and Mavs -/- BMMs 24 hr post infection. Uninfected WT set to 1.0. (D) Flow cytometry analysis for ICAM-1 expression by uninfected or M . avium 104 infected WT and Mavs -/- BMMs 24 hr post infection. (E) Flow cytometry analysis for ICAM-1 surface expression on AMs isolated from uninfected or M . avium -infected WT and Mavs -/- mice 3 weeks post infection. (F) qRT-PCR for ICAM-1 in M . avium -infected WT BMMs (24 hr post infection) that were pretreated with control, RIG-I, TBK1, IRF3 or IRF7 siRNA. Control uninfected set to 1.0 Data shown in (C and F) are the mean ±SD (n = 3 wells per group), and all data are representative of two (A and B) or three (C-F) independent experiments. In qRT-PCR, GAPDH served as loading control and data was expressed as fold change relative to uninfected WT (C), or control uninfected (F) cells. n.s., not statistically significant and **P
Figure Legend Snippet: M . avium induces ICAM-1 production in macrophages through a RIG-I/MAVS/TBK1-dependent pathway. (A) Cytokine array analyzing indicated serum proteins in WT and Mavs -/- mice 3 weeks post M . avium intratracheal infection using a dose of 1x10 7 CFU/mouse. (B) Densitometry of serum proteins highlighted in (A). (C) qRT-PCR analysis of designated transcripts in uninfected or M . avium 104 infected WT and Mavs -/- BMMs 24 hr post infection. Uninfected WT set to 1.0. (D) Flow cytometry analysis for ICAM-1 expression by uninfected or M . avium 104 infected WT and Mavs -/- BMMs 24 hr post infection. (E) Flow cytometry analysis for ICAM-1 surface expression on AMs isolated from uninfected or M . avium -infected WT and Mavs -/- mice 3 weeks post infection. (F) qRT-PCR for ICAM-1 in M . avium -infected WT BMMs (24 hr post infection) that were pretreated with control, RIG-I, TBK1, IRF3 or IRF7 siRNA. Control uninfected set to 1.0 Data shown in (C and F) are the mean ±SD (n = 3 wells per group), and all data are representative of two (A and B) or three (C-F) independent experiments. In qRT-PCR, GAPDH served as loading control and data was expressed as fold change relative to uninfected WT (C), or control uninfected (F) cells. n.s., not statistically significant and **P

Techniques Used: Mouse Assay, Infection, Quantitative RT-PCR, Flow Cytometry, Expressing, Affinity Magnetic Separation, Isolation

Increased ETV5 protein stability through RIG-I/MAVS/TBK1 activation is required for M . avium -induced ICAM-1 expression in macrophages. (A) Transcription factors and recognition sites on the promoter of mouse and human ICAM-1 gene. (B) Western blot analysis for transcription factors (A) in the nuclear fraction (Nucleus) and WCL of WT and Mavs -/- BMMs uninfected or infected with M . avium for 24 hr. (C) Western blot analysis for ETV5 in BMMs 24 hr after M . avium -infection in cells that were pretreated with control, RIG-I or TBK1 siRNA. (D) qRT-PCR for ETV5 in WT and Mavs -/- BMMs uninfected or infected with M . avium for 24 hr. Uninfected WT set to 1.0. (E) Western blot for ETV5 in nuclear fraction (Nucleus) and WCL of Mavs -/- BMMs infected with M . avium for 24 hr and treated with different concentrations of MLN4924 at. (F) qRT-PCR for ETV5 in Mavs -/- BMMs following a 24 hr infection with M . avium 104 treated with MLN4924 at various concentrations. Uninfected WT set to 1.0. (G) Western blot analysis for ETV5 in BMMs infected for 24 hr with M. avium 104. Cells were pretreated with negative control or EVT5 siRNA for 48 hr prior to infection. (H) qRT-PCR for ICAM-1 production in WT and Mavs -/- BMMs pretreated with control or ETV5 siRNA, and then left uninfected or infected with M . avium for 24 hr. Uninfected WT set to 1.0. (I) Similar to (H), but flow cytometry analysis for ICAM-1. (J) Similar to (F), but qRT-PCR for ICAM-1. (K) Flow cytometry analysis for ICAM-1 in Mavs -/- BMMs infected with M . avium for 24 hr in the presence of MLN4924 (0.5 μM). Data shown in (D, F, H and J) are the mean ±SD (n = 3 wells per group), and all data are representative of three independent experiments. In qRT-PCR, GAPDH served as loading control and data was expressed as fold change relative to uninfected WT cells or control siRNA-treated uninfected WT cells. n.s., not statistically significant and *P
Figure Legend Snippet: Increased ETV5 protein stability through RIG-I/MAVS/TBK1 activation is required for M . avium -induced ICAM-1 expression in macrophages. (A) Transcription factors and recognition sites on the promoter of mouse and human ICAM-1 gene. (B) Western blot analysis for transcription factors (A) in the nuclear fraction (Nucleus) and WCL of WT and Mavs -/- BMMs uninfected or infected with M . avium for 24 hr. (C) Western blot analysis for ETV5 in BMMs 24 hr after M . avium -infection in cells that were pretreated with control, RIG-I or TBK1 siRNA. (D) qRT-PCR for ETV5 in WT and Mavs -/- BMMs uninfected or infected with M . avium for 24 hr. Uninfected WT set to 1.0. (E) Western blot for ETV5 in nuclear fraction (Nucleus) and WCL of Mavs -/- BMMs infected with M . avium for 24 hr and treated with different concentrations of MLN4924 at. (F) qRT-PCR for ETV5 in Mavs -/- BMMs following a 24 hr infection with M . avium 104 treated with MLN4924 at various concentrations. Uninfected WT set to 1.0. (G) Western blot analysis for ETV5 in BMMs infected for 24 hr with M. avium 104. Cells were pretreated with negative control or EVT5 siRNA for 48 hr prior to infection. (H) qRT-PCR for ICAM-1 production in WT and Mavs -/- BMMs pretreated with control or ETV5 siRNA, and then left uninfected or infected with M . avium for 24 hr. Uninfected WT set to 1.0. (I) Similar to (H), but flow cytometry analysis for ICAM-1. (J) Similar to (F), but qRT-PCR for ICAM-1. (K) Flow cytometry analysis for ICAM-1 in Mavs -/- BMMs infected with M . avium for 24 hr in the presence of MLN4924 (0.5 μM). Data shown in (D, F, H and J) are the mean ±SD (n = 3 wells per group), and all data are representative of three independent experiments. In qRT-PCR, GAPDH served as loading control and data was expressed as fold change relative to uninfected WT cells or control siRNA-treated uninfected WT cells. n.s., not statistically significant and *P

Techniques Used: Activation Assay, Expressing, Western Blot, Infection, Quantitative RT-PCR, Negative Control, Flow Cytometry

ICAM-1 production in host cells is essential for CD4 + T cell-mediated M . avium killing in macrophage and mice. (A) Confocal microscopy measuring the cell-cell interaction between M . avium (tdTomato)-infected WT, Mavs -/- or ICAM-1 -/- BMMs and CD4 + T cells (Alexa Fluor 488) isolated from M . avium -infected WT mice +/- MLN4924 (0.5 μM). Images were taken 24 hr after BMM and CD4 + T cell coculture. Scale bar, 20 μm. For quantification, all data was normalized to one biological repeat in WT group. (B) M . avium burden in Mavs -/- BMMs pretreated with MLN4924 (0.5 μΜ) and then cocultured with CD4 + T cells isolated from M . avium -infected WT or Mavs -/- mice. (C) M . avium burden in WT and ICAM-1 -/- BMMs cocultured with CD4 + T cells isolated from M . avium -infected WT mice. (D) IFN-γ concentration in the culture supernatant of uninfected or M . avium -infected WT and ICAM-1 -/- BMMs cocultured for 72 hr with CD4 + T cells isolated from M . avium -infected WT mice. (E) ROS measured over time in M . avium -infected WT and ICAM-1 -/- BMMs +/- coculture with CD4 + T cells isolated from M . avium -infected WT mice. (F) Western blot for iNOS in M . avium -infected WT and ICAM-1 -/- BMMs +/- coculture for 24 hr with CD4 + T cells isolated from M . avium -infected WT mice. Quantitation of band pixel intensity shown in graph. (G) Gen R and Gen S M . avium burden in WT mouse lung at day 1, 3 and 6 after M . avium -infected AM adoptive transfer. WT AM and ICAM-1 -/- AMs were isolated from WT and ICAM-1 -/- naïve mice, respectively. Data in (A-E) are the mean ±SD (n = 3 independent infections per group), and data shown are representative of at least three (A-F) or combination of two (G, n = 3 per group each experiment) independent experiments. The ratio between BMMs and CD4 + T cells was 1:2. n.s., not statistically significant, *P
Figure Legend Snippet: ICAM-1 production in host cells is essential for CD4 + T cell-mediated M . avium killing in macrophage and mice. (A) Confocal microscopy measuring the cell-cell interaction between M . avium (tdTomato)-infected WT, Mavs -/- or ICAM-1 -/- BMMs and CD4 + T cells (Alexa Fluor 488) isolated from M . avium -infected WT mice +/- MLN4924 (0.5 μM). Images were taken 24 hr after BMM and CD4 + T cell coculture. Scale bar, 20 μm. For quantification, all data was normalized to one biological repeat in WT group. (B) M . avium burden in Mavs -/- BMMs pretreated with MLN4924 (0.5 μΜ) and then cocultured with CD4 + T cells isolated from M . avium -infected WT or Mavs -/- mice. (C) M . avium burden in WT and ICAM-1 -/- BMMs cocultured with CD4 + T cells isolated from M . avium -infected WT mice. (D) IFN-γ concentration in the culture supernatant of uninfected or M . avium -infected WT and ICAM-1 -/- BMMs cocultured for 72 hr with CD4 + T cells isolated from M . avium -infected WT mice. (E) ROS measured over time in M . avium -infected WT and ICAM-1 -/- BMMs +/- coculture with CD4 + T cells isolated from M . avium -infected WT mice. (F) Western blot for iNOS in M . avium -infected WT and ICAM-1 -/- BMMs +/- coculture for 24 hr with CD4 + T cells isolated from M . avium -infected WT mice. Quantitation of band pixel intensity shown in graph. (G) Gen R and Gen S M . avium burden in WT mouse lung at day 1, 3 and 6 after M . avium -infected AM adoptive transfer. WT AM and ICAM-1 -/- AMs were isolated from WT and ICAM-1 -/- naïve mice, respectively. Data in (A-E) are the mean ±SD (n = 3 independent infections per group), and data shown are representative of at least three (A-F) or combination of two (G, n = 3 per group each experiment) independent experiments. The ratio between BMMs and CD4 + T cells was 1:2. n.s., not statistically significant, *P

Techniques Used: Mouse Assay, Confocal Microscopy, Infection, Isolation, Concentration Assay, Western Blot, Quantitation Assay, Adoptive Transfer Assay, Affinity Magnetic Separation

3) Product Images from "Osteogenic protein-1 reduces intercellular adhesion molecule-1 messenger RNA expression, infarct size and TUNEL-positive cardiomyocytes in ischemia/reperfusion rat hearts"

Article Title: Osteogenic protein-1 reduces intercellular adhesion molecule-1 messenger RNA expression, infarct size and TUNEL-positive cardiomyocytes in ischemia/reperfusion rat hearts

Journal: Experimental & Clinical Cardiology

doi:

A Results from Northern blotting for intracellular adhesion molecule-1 (ICAM-1) messenger RNA (mRNA) expression in a preligation heart and hearts that underwent ischemia/reperfusion with osteogenic protein-1 or vehicle treatment. Lane 1 – preligation heart; lanes 2, 3 and 4 – 4 h, one-day and seven-day hearts with osteogenic protein-1 treatment, respectively; lanes 5, 6 and 7 – 4 h, one-day and seven-day ischemia/reperfusion hearts treated with the vehicle, respectively. Expression of ICAM-1 mRNA in ischemia/reperfusion hearts was lower in osteogenic protein-1-treated rats than in vehicle-treated rats. The hybridized band obtained from a preligation heart (lane 1) was used to standardize the expression of ICAM-1 mRNA from both osteogenic protein-1- and vehicle-treated ischemia/reperfusion hearts. B Summarized results of osteogenic protein-1 mRNA expression relative to loaded 28S ribsosmal RNA stained with ethidium bromide. The data are expressed relative to the values in preligation hearts, taken as 1.00. Significantly reduced expression of ICAM-1 mRNA in ischemia/reperfusion hearts was seen in the osteogenic protein-1-treated rats compared with the untreated rats. Data are expressed as mean ± SEM of three hearts from each group
Figure Legend Snippet: A Results from Northern blotting for intracellular adhesion molecule-1 (ICAM-1) messenger RNA (mRNA) expression in a preligation heart and hearts that underwent ischemia/reperfusion with osteogenic protein-1 or vehicle treatment. Lane 1 – preligation heart; lanes 2, 3 and 4 – 4 h, one-day and seven-day hearts with osteogenic protein-1 treatment, respectively; lanes 5, 6 and 7 – 4 h, one-day and seven-day ischemia/reperfusion hearts treated with the vehicle, respectively. Expression of ICAM-1 mRNA in ischemia/reperfusion hearts was lower in osteogenic protein-1-treated rats than in vehicle-treated rats. The hybridized band obtained from a preligation heart (lane 1) was used to standardize the expression of ICAM-1 mRNA from both osteogenic protein-1- and vehicle-treated ischemia/reperfusion hearts. B Summarized results of osteogenic protein-1 mRNA expression relative to loaded 28S ribsosmal RNA stained with ethidium bromide. The data are expressed relative to the values in preligation hearts, taken as 1.00. Significantly reduced expression of ICAM-1 mRNA in ischemia/reperfusion hearts was seen in the osteogenic protein-1-treated rats compared with the untreated rats. Data are expressed as mean ± SEM of three hearts from each group

Techniques Used: Northern Blot, Expressing, Staining

Related Articles

Mutagenesis:

Article Title: Efficient gene replacement and direct hyphal transformation in Sclerotinia sclerotiorum
Article Snippet: .. For Southern blot analysis, total genomic DNA was extracted from ssku80 mutant strain 20 and the wild type, digested with either Bam HI or Eco RV and transferred to a Hybond‐XL (Amersham Biosciences, UK) nylon membrane. .. Hybridization was performed at 42 °C under low‐stringency conditions in the presence of ULTRAhyb solution (Ambion, Austin, TX).

Electrophoresis:

Article Title: Regulation of RNA polymerase III transcription during transformation of human IMR90 fibroblasts with defined genetic elements
Article Snippet: .. 8 µg of total RNA was separated by electrophoresis and blotted onto Hybond-XL nylon membrane (GE Healthcare, RPN2020S). .. The membrane was incubated in 1x Church buffer (0.25 M NaH2 PO4 ; 1 mM EDTA; 7% SDS; 20 mg/ml salmon sperm DNA; 0.5% BSA) for 2 hrs at 28°C and the denatured radioactive probe was added for incubation overnight.

Agarose Gel Electrophoresis:

Article Title: Modeling cancer genomic data in yeast reveals selection against ATM function during tumorigenesis
Article Snippet: .. Genomic DNA was either Pst I-or Xho I-digested (as specified in Figure legends), run on 1.3% agarose gel and transferred on an Amersham Hybond-XL membrane (GE Healthcare) and detected by Southern blot with a 32 P-labeled telomere-specific probe (5′-TGTGGTGTGTGGGTGTGGTGT-3′) as described [ ]. .. Rad50 overexpression and purification from yeast cells The yeast galactose inducible expression plasmid, pR50.1 (2μ, GAL-PGK-RAD50, leu2-d), was a gift from Patrick Sung [ ].

Article Title: rahu is a mutant allele of Dnmt3c, encoding a DNA methyltransferase homolog required for meiosis and transposon repression in the mouse male germline
Article Snippet: .. ~250 ng of digested DNA was electrophoresed on a 0.9% agarose gel and transferred as described [ ] to an Amersham Hybond-XL membrane (GE Healthcare). .. The L1 5′UTR probe has been described elsewhere [ ] and corresponds to nucleotides 515–1628 of the L1 sequence (GenBank accession number M13002).

Southern Blot:

Article Title: Efficient gene replacement and direct hyphal transformation in Sclerotinia sclerotiorum
Article Snippet: .. For Southern blot analysis, total genomic DNA was extracted from ssku80 mutant strain 20 and the wild type, digested with either Bam HI or Eco RV and transferred to a Hybond‐XL (Amersham Biosciences, UK) nylon membrane. .. Hybridization was performed at 42 °C under low‐stringency conditions in the presence of ULTRAhyb solution (Ambion, Austin, TX).

Article Title: Preferential Repair of the Transcribed DNA Strand in Plants
Article Snippet: .. Samples (at least 1 μg of DNA/lane) were electrophoresed on a 0.5% alkaline gel (9.5 cm) in 1 mM EDTA and 30 mM NaOH at 22 V for around 16 h. Southern blotting, hybridization and washes were performed as described (Spivak and Hanawalt Philip, ) with the exception that Hybond-N+ or Hybond-XL nylon membranes (Amersham Pharmacia, UK) were used. ..

Article Title: Modeling cancer genomic data in yeast reveals selection against ATM function during tumorigenesis
Article Snippet: .. Genomic DNA was either Pst I-or Xho I-digested (as specified in Figure legends), run on 1.3% agarose gel and transferred on an Amersham Hybond-XL membrane (GE Healthcare) and detected by Southern blot with a 32 P-labeled telomere-specific probe (5′-TGTGGTGTGTGGGTGTGGTGT-3′) as described [ ]. .. Rad50 overexpression and purification from yeast cells The yeast galactose inducible expression plasmid, pR50.1 (2μ, GAL-PGK-RAD50, leu2-d), was a gift from Patrick Sung [ ].

Hybridization:

Article Title: Preferential Repair of the Transcribed DNA Strand in Plants
Article Snippet: .. Samples (at least 1 μg of DNA/lane) were electrophoresed on a 0.5% alkaline gel (9.5 cm) in 1 mM EDTA and 30 mM NaOH at 22 V for around 16 h. Southern blotting, hybridization and washes were performed as described (Spivak and Hanawalt Philip, ) with the exception that Hybond-N+ or Hybond-XL nylon membranes (Amersham Pharmacia, UK) were used. ..

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    GE Healthcare recombinant fc tagged icam 1
    Plasma levels of IgG with specificity for P. falciparum DBLβ domains. Samples were obtained from 79 Ghanaian children with either severe (▲) or nonsevere P. falciparum malaria. (A) Levels (ELISA units [EU]) in plasma from individual children (columns) of IgG antibodies specific for individual group A DBLβ domains (rows) containing (DBLβ_motif; M1 to M12, M14, and M15) or not containing (DBLβ_nonmotif; N20 to N32; lower half) the <t>ICAM-1-binding</t> motif identified by Lennartz et al. ( 33 ). Shading indicates IgG level score: black, score of 4 ( > 100 EU); dark gray, score of 3 (76 to 100 EU); gray, score of 2 (51 to 75 EU); light gray, score of 1 (26 to 50 EU); white, score of 0 (0 to 25 EU). The DBLβ domain numbers correspond to the numbers in Table 3 . Danish controls ( n = 25) did not react with any of the domains (data not shown). (B) The means of IgG level scores (defined as described for panel A) of individual DBLβ domains that contain (motif; ○) or do not contain (nonmotif; ●) the ICAM-1-binding motif. Error bars indicate 95% confidence intervals. DBLβ domain numbering is as described for panel A. (C) The means of IgG level scores (defined as described for panel A) of individual children for IgG specific for DBLβ_motif (○) and DBLβ_nonmotif (●) domains. The statistical significance (Mann-Whitney rank sum test) of pairwise comparisons is shown along the top of the panel.
    Recombinant Fc Tagged Icam 1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant fc tagged icam 1/product/GE Healthcare
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    Price from $9.99 to $1999.99
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    GE Healthcare icam 1
    Vpu prevents <t>ICAM-1</t> and ICAM-3 upregulation on the surface of primary CD4 + T cells by a process that depends on its TMD and dual-serine motif. Primary human CD4 + T cells were infected with GFP-marked NL4.3 HIV-1 either lacking Vpu (ΔVpu) or expressing
    Icam 1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icam 1/product/GE Healthcare
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    GE Healthcare recombinant protein purification recombinant human icam 5 d1 4 fc
    sICAM-5 promotes dendritic filopodia elongation. 9-DIV EGFP-transfected WT and <t>ICAM-5</t> −/− hippocampal neurons were incubated with 10 μg/ml soluble recombinant ICAM-5 <t>D1-4-Fc</t> protein or control mIgG for 72 h. The neurons were then double stained for ICAM-5 (A, red) and MAP-2 (A, blue). Compared with mIgG control protein, sICAM-5 D1-4-Fc protein induced significantly more filopodia from WT neurons, but not from ICAM-5 −/− neurons (B and C). The filopodial length of WT neurons, but not ICAM-5 −/− neurons, also significantly increased in the presence of sICAM-5 D1-4-Fc protein (D). The experiment was repeated three times with similar results. Error bars indicate mean ± SD. *, P
    Recombinant Protein Purification Recombinant Human Icam 5 D1 4 Fc, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Plasma levels of IgG with specificity for P. falciparum DBLβ domains. Samples were obtained from 79 Ghanaian children with either severe (▲) or nonsevere P. falciparum malaria. (A) Levels (ELISA units [EU]) in plasma from individual children (columns) of IgG antibodies specific for individual group A DBLβ domains (rows) containing (DBLβ_motif; M1 to M12, M14, and M15) or not containing (DBLβ_nonmotif; N20 to N32; lower half) the ICAM-1-binding motif identified by Lennartz et al. ( 33 ). Shading indicates IgG level score: black, score of 4 ( > 100 EU); dark gray, score of 3 (76 to 100 EU); gray, score of 2 (51 to 75 EU); light gray, score of 1 (26 to 50 EU); white, score of 0 (0 to 25 EU). The DBLβ domain numbers correspond to the numbers in Table 3 . Danish controls ( n = 25) did not react with any of the domains (data not shown). (B) The means of IgG level scores (defined as described for panel A) of individual DBLβ domains that contain (motif; ○) or do not contain (nonmotif; ●) the ICAM-1-binding motif. Error bars indicate 95% confidence intervals. DBLβ domain numbering is as described for panel A. (C) The means of IgG level scores (defined as described for panel A) of individual children for IgG specific for DBLβ_motif (○) and DBLβ_nonmotif (●) domains. The statistical significance (Mann-Whitney rank sum test) of pairwise comparisons is shown along the top of the panel.

    Journal: Infection and Immunity

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria

    doi: 10.1128/IAI.00622-17

    Figure Lengend Snippet: Plasma levels of IgG with specificity for P. falciparum DBLβ domains. Samples were obtained from 79 Ghanaian children with either severe (▲) or nonsevere P. falciparum malaria. (A) Levels (ELISA units [EU]) in plasma from individual children (columns) of IgG antibodies specific for individual group A DBLβ domains (rows) containing (DBLβ_motif; M1 to M12, M14, and M15) or not containing (DBLβ_nonmotif; N20 to N32; lower half) the ICAM-1-binding motif identified by Lennartz et al. ( 33 ). Shading indicates IgG level score: black, score of 4 ( > 100 EU); dark gray, score of 3 (76 to 100 EU); gray, score of 2 (51 to 75 EU); light gray, score of 1 (26 to 50 EU); white, score of 0 (0 to 25 EU). The DBLβ domain numbers correspond to the numbers in Table 3 . Danish controls ( n = 25) did not react with any of the domains (data not shown). (B) The means of IgG level scores (defined as described for panel A) of individual DBLβ domains that contain (motif; ○) or do not contain (nonmotif; ●) the ICAM-1-binding motif. Error bars indicate 95% confidence intervals. DBLβ domain numbering is as described for panel A. (C) The means of IgG level scores (defined as described for panel A) of individual children for IgG specific for DBLβ_motif (○) and DBLβ_nonmotif (●) domains. The statistical significance (Mann-Whitney rank sum test) of pairwise comparisons is shown along the top of the panel.

    Article Snippet: Recombinant Fc-tagged ICAM-1 was expressed in HEK293 cells and purified on a HiTrap Protein G HP column ( http://www3.gehealthcare.dk/ ) as described previously ( ).

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, MANN-WHITNEY

    Ability of DBLβ_motif-specific IgG to inhibit binding of ICAM-1 to DBLβ domains. (A) Inhibition of ICAM-1 binding by pooled immune plasma from 10 of the study children (pool). ***, P F

    Journal: Infection and Immunity

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria

    doi: 10.1128/IAI.00622-17

    Figure Lengend Snippet: Ability of DBLβ_motif-specific IgG to inhibit binding of ICAM-1 to DBLβ domains. (A) Inhibition of ICAM-1 binding by pooled immune plasma from 10 of the study children (pool). ***, P F

    Article Snippet: Recombinant Fc-tagged ICAM-1 was expressed in HEK293 cells and purified on a HiTrap Protein G HP column ( http://www3.gehealthcare.dk/ ) as described previously ( ).

    Techniques: Binding Assay, Inhibition

    ICAM-1-specific adhesion of erythrocytes infected by patient P. falciparum isolates and inhibition of ICAM-1-adhering IEs by M6pep/M9pep-specific antibody. (A) Adhesion of 33 patient isolates to ICAM-1 under physiologic flow; (B) antibody-mediated inhibition ( > 25%) of ICAM-1-specific IE adhesion among the 22 patient isolates adhering (≥10 adherent IEs/mm 2 ) to ICAM-1 under physiologic flow. The isolates were tested in one experiment with five technical replicates. Fewer than 0.25 IEs/mm 2 bound to uncoated channels. Isolates from patients with uncomplicated malaria (○), cerebral malaria (▲), and noncerebral severe disease (●) are indicated in both panels.

    Journal: Infection and Immunity

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria

    doi: 10.1128/IAI.00622-17

    Figure Lengend Snippet: ICAM-1-specific adhesion of erythrocytes infected by patient P. falciparum isolates and inhibition of ICAM-1-adhering IEs by M6pep/M9pep-specific antibody. (A) Adhesion of 33 patient isolates to ICAM-1 under physiologic flow; (B) antibody-mediated inhibition ( > 25%) of ICAM-1-specific IE adhesion among the 22 patient isolates adhering (≥10 adherent IEs/mm 2 ) to ICAM-1 under physiologic flow. The isolates were tested in one experiment with five technical replicates. Fewer than 0.25 IEs/mm 2 bound to uncoated channels. Isolates from patients with uncomplicated malaria (○), cerebral malaria (▲), and noncerebral severe disease (●) are indicated in both panels.

    Article Snippet: Recombinant Fc-tagged ICAM-1 was expressed in HEK293 cells and purified on a HiTrap Protein G HP column ( http://www3.gehealthcare.dk/ ) as described previously ( ).

    Techniques: Infection, Inhibition, Flow Cytometry

    DBLβ-specific antibody-mediated inhibition of adhesion of IEs to ICAM-1 under physiologic shear stress, relative to that of control without antibody (none). (A) IEs expressing PFD1235w; (B) IEs expressing HB3VAR03; (C) IEs expressing IT4VAR13. The specificities of the DBLβ antibodies correspond to the data in Table 3 . The antiserum marked with an asterisk was affinity purified on a peptide (M6pep) representing the binding motif in PFD1235w DBLβ_D4 prior to assaying. An ICAM-1-specific neutralizing antibody (ICAM-1) and an irrelevant rat anti-IgG (Sigma-Aldrich) were included as positive and negative controls, respectively. Fewer than 0.25 IEs/mm 2 bound to uncoated channels. Means (bars) and standard deviations (error bars) of the results of at least three independent experiments in triplicate are shown. Statistically significant reductions relative to adhesion in the absence of antibody (−) are indicated above the bars (**, P F

    Journal: Infection and Immunity

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria

    doi: 10.1128/IAI.00622-17

    Figure Lengend Snippet: DBLβ-specific antibody-mediated inhibition of adhesion of IEs to ICAM-1 under physiologic shear stress, relative to that of control without antibody (none). (A) IEs expressing PFD1235w; (B) IEs expressing HB3VAR03; (C) IEs expressing IT4VAR13. The specificities of the DBLβ antibodies correspond to the data in Table 3 . The antiserum marked with an asterisk was affinity purified on a peptide (M6pep) representing the binding motif in PFD1235w DBLβ_D4 prior to assaying. An ICAM-1-specific neutralizing antibody (ICAM-1) and an irrelevant rat anti-IgG (Sigma-Aldrich) were included as positive and negative controls, respectively. Fewer than 0.25 IEs/mm 2 bound to uncoated channels. Means (bars) and standard deviations (error bars) of the results of at least three independent experiments in triplicate are shown. Statistically significant reductions relative to adhesion in the absence of antibody (−) are indicated above the bars (**, P F

    Article Snippet: Recombinant Fc-tagged ICAM-1 was expressed in HEK293 cells and purified on a HiTrap Protein G HP column ( http://www3.gehealthcare.dk/ ) as described previously ( ).

    Techniques: Inhibition, Expressing, Affinity Purification, Binding Assay

    Sequence logo showing the ICAM-1-binding motif (as defined in reference 33 ) of DBLβ domains 1 to 15 used in the present study ( Table 3 ). Residues that are critical for the direct interaction with ICAM-1 (red triangles) or for the architecture of the ICAM-1 binding (white triangles) are indicated. The group A PfEMP1 ICAM-1-binding motif was identified by Lennartz et al. ( 33 ).

    Journal: Infection and Immunity

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria

    doi: 10.1128/IAI.00622-17

    Figure Lengend Snippet: Sequence logo showing the ICAM-1-binding motif (as defined in reference 33 ) of DBLβ domains 1 to 15 used in the present study ( Table 3 ). Residues that are critical for the direct interaction with ICAM-1 (red triangles) or for the architecture of the ICAM-1 binding (white triangles) are indicated. The group A PfEMP1 ICAM-1-binding motif was identified by Lennartz et al. ( 33 ).

    Article Snippet: Recombinant Fc-tagged ICAM-1 was expressed in HEK293 cells and purified on a HiTrap Protein G HP column ( http://www3.gehealthcare.dk/ ) as described previously ( ).

    Techniques: Sequencing, Binding Assay

    Ability of rat antisera to the ICAM-1-binding motif in DBLβ_motif domains to inhibit binding of recombinant DBLβ domains to ICAM-1. (A) Antisera from rats immunized with M6pep or with both M6pep and M9pep tested against recombinant DBLβ_motif domains (M2 to M7, M9, and M11 to M13) and DBLβ_nonmotif domains (N27 and N33). Shading, DBLβ domain numbers, and antiserum specificities are as described in the legend to Fig. 3B . (B to D) Inhibition by the same antisera of ICAM-1-specific adhesion of PFD1235w-positive IEs (B), HB3VAR03-positive IEs (C), and IT4VAR13-positive IEs (D) under physiologic shear stress. The statistical significance of the reductions is indicated as described in the legend to Fig. 4 . Three independent experiments were done (with three technical replicates in each). Fewer than 0.25 IEs/mm 2 bound to uncoated channels were observed. (E) Immunofluorescence of representative IEs with surface expression of PFD1235w (top row), HB3VAR03 (center row), and IT4VAR13 (bottom row) and labeled by sera from rats immunized with M6pep only (left column) or with both M6pep and M9pep (center column), or by a rat antiserum to N27 (right column). See Table S2 in the supplemental material for raw data.

    Journal: Infection and Immunity

    Article Title: Natural and Vaccine-Induced Acquisition of Cross-Reactive IgG-Inhibiting ICAM-1-Specific Binding of a Plasmodium falciparum PfEMP1 Subtype Associated Specifically with Cerebral Malaria

    doi: 10.1128/IAI.00622-17

    Figure Lengend Snippet: Ability of rat antisera to the ICAM-1-binding motif in DBLβ_motif domains to inhibit binding of recombinant DBLβ domains to ICAM-1. (A) Antisera from rats immunized with M6pep or with both M6pep and M9pep tested against recombinant DBLβ_motif domains (M2 to M7, M9, and M11 to M13) and DBLβ_nonmotif domains (N27 and N33). Shading, DBLβ domain numbers, and antiserum specificities are as described in the legend to Fig. 3B . (B to D) Inhibition by the same antisera of ICAM-1-specific adhesion of PFD1235w-positive IEs (B), HB3VAR03-positive IEs (C), and IT4VAR13-positive IEs (D) under physiologic shear stress. The statistical significance of the reductions is indicated as described in the legend to Fig. 4 . Three independent experiments were done (with three technical replicates in each). Fewer than 0.25 IEs/mm 2 bound to uncoated channels were observed. (E) Immunofluorescence of representative IEs with surface expression of PFD1235w (top row), HB3VAR03 (center row), and IT4VAR13 (bottom row) and labeled by sera from rats immunized with M6pep only (left column) or with both M6pep and M9pep (center column), or by a rat antiserum to N27 (right column). See Table S2 in the supplemental material for raw data.

    Article Snippet: Recombinant Fc-tagged ICAM-1 was expressed in HEK293 cells and purified on a HiTrap Protein G HP column ( http://www3.gehealthcare.dk/ ) as described previously ( ).

    Techniques: Binding Assay, Recombinant, Inhibition, Immunofluorescence, Expressing, Labeling

    Vpu prevents ICAM-1 and ICAM-3 upregulation on the surface of primary CD4 + T cells by a process that depends on its TMD and dual-serine motif. Primary human CD4 + T cells were infected with GFP-marked NL4.3 HIV-1 either lacking Vpu (ΔVpu) or expressing

    Journal: Journal of Virology

    Article Title: HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells

    doi: 10.1128/JVI.02442-16

    Figure Lengend Snippet: Vpu prevents ICAM-1 and ICAM-3 upregulation on the surface of primary CD4 + T cells by a process that depends on its TMD and dual-serine motif. Primary human CD4 + T cells were infected with GFP-marked NL4.3 HIV-1 either lacking Vpu (ΔVpu) or expressing

    Article Snippet: ICAM-1 was immunoprecipitated using an excess of goat anti-ICAM-1 Abs, and radiolabeled immunoprecipitates were separated via SDS-PAGE and analyzed using a Storm 860 molecular imager (Molecular Dynamics, GE Healthcare) as described previously ( ).

    Techniques: Infection, Expressing

    ICAM-1 is upregulated at the transcriptional level in response to HIV-1 infection and downregulated posttranscriptionally by Vpu. HeLa cells were infected with VSV-G-pseudotyped GFP-marked wild-type NL4.3 HIV-1 (WT) or HIV-1 lacking Vpu (ΔVpu)

    Journal: Journal of Virology

    Article Title: HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells

    doi: 10.1128/JVI.02442-16

    Figure Lengend Snippet: ICAM-1 is upregulated at the transcriptional level in response to HIV-1 infection and downregulated posttranscriptionally by Vpu. HeLa cells were infected with VSV-G-pseudotyped GFP-marked wild-type NL4.3 HIV-1 (WT) or HIV-1 lacking Vpu (ΔVpu)

    Article Snippet: ICAM-1 was immunoprecipitated using an excess of goat anti-ICAM-1 Abs, and radiolabeled immunoprecipitates were separated via SDS-PAGE and analyzed using a Storm 860 molecular imager (Molecular Dynamics, GE Healthcare) as described previously ( ).

    Techniques: Infection

    Vpu-induced ICAM-1 downregulation prevents NK cell conjugation to HIV-1-infected CD4 + T cells. (A and B) Sorted GFP + WT or Vpu-deficient (ΔVpu) HIV-1-infected primary CD4 + T cells were cocultured with eFluor 670-labeled autologous NK cells at

    Journal: Journal of Virology

    Article Title: HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells

    doi: 10.1128/JVI.02442-16

    Figure Lengend Snippet: Vpu-induced ICAM-1 downregulation prevents NK cell conjugation to HIV-1-infected CD4 + T cells. (A and B) Sorted GFP + WT or Vpu-deficient (ΔVpu) HIV-1-infected primary CD4 + T cells were cocultured with eFluor 670-labeled autologous NK cells at

    Article Snippet: ICAM-1 was immunoprecipitated using an excess of goat anti-ICAM-1 Abs, and radiolabeled immunoprecipitates were separated via SDS-PAGE and analyzed using a Storm 860 molecular imager (Molecular Dynamics, GE Healthcare) as described previously ( ).

    Techniques: Conjugation Assay, Infection, Labeling

    Vpu dual-serine-motif-dependent surface ICAM-1 downregulation is dissociable from β-TrCP-1-mediated ICAM-1 degradation. (A) Expression of β-TrCP-1, β-TrCP-2, and BST2 mRNA transcripts was evaluated by RT-qPCR in HeLa cells expressing

    Journal: Journal of Virology

    Article Title: HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells

    doi: 10.1128/JVI.02442-16

    Figure Lengend Snippet: Vpu dual-serine-motif-dependent surface ICAM-1 downregulation is dissociable from β-TrCP-1-mediated ICAM-1 degradation. (A) Expression of β-TrCP-1, β-TrCP-2, and BST2 mRNA transcripts was evaluated by RT-qPCR in HeLa cells expressing

    Article Snippet: ICAM-1 was immunoprecipitated using an excess of goat anti-ICAM-1 Abs, and radiolabeled immunoprecipitates were separated via SDS-PAGE and analyzed using a Storm 860 molecular imager (Molecular Dynamics, GE Healthcare) as described previously ( ).

    Techniques: Expressing, Quantitative RT-PCR

    Vpu is sufficient to deplete endogenous ICAM-1 in a TMD- and dual-serine-motif-dependent manner. HeLa cells were transfected to express wild-type NL4.3 Vpu (pVpuWT), a mutant Vpu lacking the dual-serine motif (pVpuS52,56N), a mutant Vpu containing a randomized

    Journal: Journal of Virology

    Article Title: HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells

    doi: 10.1128/JVI.02442-16

    Figure Lengend Snippet: Vpu is sufficient to deplete endogenous ICAM-1 in a TMD- and dual-serine-motif-dependent manner. HeLa cells were transfected to express wild-type NL4.3 Vpu (pVpuWT), a mutant Vpu lacking the dual-serine motif (pVpuS52,56N), a mutant Vpu containing a randomized

    Article Snippet: ICAM-1 was immunoprecipitated using an excess of goat anti-ICAM-1 Abs, and radiolabeled immunoprecipitates were separated via SDS-PAGE and analyzed using a Storm 860 molecular imager (Molecular Dynamics, GE Healthcare) as described previously ( ).

    Techniques: Transfection, Mutagenesis

    Vpu lowers viral infectivity by decreasing the amount of mature ICAM-1 packaged into HIV-1 virions. (A) HEK 293T cells were cotransfected with plasmids encoding ICAM-1 (pICAM-1) or TfR (pTfR), along with GFP-marked plasmids expressing wild-type NL4.3

    Journal: Journal of Virology

    Article Title: HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells

    doi: 10.1128/JVI.02442-16

    Figure Lengend Snippet: Vpu lowers viral infectivity by decreasing the amount of mature ICAM-1 packaged into HIV-1 virions. (A) HEK 293T cells were cotransfected with plasmids encoding ICAM-1 (pICAM-1) or TfR (pTfR), along with GFP-marked plasmids expressing wild-type NL4.3

    Article Snippet: ICAM-1 was immunoprecipitated using an excess of goat anti-ICAM-1 Abs, and radiolabeled immunoprecipitates were separated via SDS-PAGE and analyzed using a Storm 860 molecular imager (Molecular Dynamics, GE Healthcare) as described previously ( ).

    Techniques: Infection, Expressing

    Vpu interacts with ICAM-1 and induces its turnover in a proteasome-dependent manner. (A) HeLa cells were infected with VSV-G-pseudotyped GFP-marked wild-type (WT); Vpu-deficient (ΔVpu); or A10/A14/A18L Vpu (A10,14,18L) NL4.3 HIV-1 for 48 h prior

    Journal: Journal of Virology

    Article Title: HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells

    doi: 10.1128/JVI.02442-16

    Figure Lengend Snippet: Vpu interacts with ICAM-1 and induces its turnover in a proteasome-dependent manner. (A) HeLa cells were infected with VSV-G-pseudotyped GFP-marked wild-type (WT); Vpu-deficient (ΔVpu); or A10/A14/A18L Vpu (A10,14,18L) NL4.3 HIV-1 for 48 h prior

    Article Snippet: ICAM-1 was immunoprecipitated using an excess of goat anti-ICAM-1 Abs, and radiolabeled immunoprecipitates were separated via SDS-PAGE and analyzed using a Storm 860 molecular imager (Molecular Dynamics, GE Healthcare) as described previously ( ).

    Techniques: Infection

    Vpu-induced ICAM-1 downregulation prevents NK cell-mediated killing of HIV-1-infected CD4 + T cells. Primary CD4 + T cells infected with GFP-marked WT, Vpu-deficient (ΔVpu), or S52/S56D Vpu (S52,56D) NL4.3 HIV-1 were cocultured with DiD-labeled

    Journal: Journal of Virology

    Article Title: HIV-1 Vpu Downmodulates ICAM-1 Expression, Resulting in Decreased Killing of Infected CD4+ T Cells by NK Cells

    doi: 10.1128/JVI.02442-16

    Figure Lengend Snippet: Vpu-induced ICAM-1 downregulation prevents NK cell-mediated killing of HIV-1-infected CD4 + T cells. Primary CD4 + T cells infected with GFP-marked WT, Vpu-deficient (ΔVpu), or S52/S56D Vpu (S52,56D) NL4.3 HIV-1 were cocultured with DiD-labeled

    Article Snippet: ICAM-1 was immunoprecipitated using an excess of goat anti-ICAM-1 Abs, and radiolabeled immunoprecipitates were separated via SDS-PAGE and analyzed using a Storm 860 molecular imager (Molecular Dynamics, GE Healthcare) as described previously ( ).

    Techniques: Infection, Labeling

    ICAM-1 induced VEC phosphorylation in wt and mutant VEC. (A) CHO-ICAM-1 cells were transfected with wt pEGFP-N’-VEC or not, grown to post-confluence and then starved. Cells were then subjected to ICAM-1 crosslinking and VEC-GFP immunoprecipitated

    Journal:

    Article Title: Phosphorylation of Vascular Endothelial Cadherin Controls Lymphocyte Emigration

    doi: 10.1242/jcs.022681

    Figure Lengend Snippet: ICAM-1 induced VEC phosphorylation in wt and mutant VEC. (A) CHO-ICAM-1 cells were transfected with wt pEGFP-N’-VEC or not, grown to post-confluence and then starved. Cells were then subjected to ICAM-1 crosslinking and VEC-GFP immunoprecipitated

    Article Snippet: Transfected CHO-ICAM-1 cells were labeled with phosphorus-32 (PBS 13; GE Healthcare; 20 MBq/ml) in phosphate-free DMEM for 3 h, ICAM-1 crosslinked, lysed and VEC-GFP precipitated using 5 μg of a polyclonal anti-GFP antibody (Abcam 290).

    Techniques: Mutagenesis, Transfection, Immunoprecipitation

    sICAM-5 promotes dendritic filopodia elongation. 9-DIV EGFP-transfected WT and ICAM-5 −/− hippocampal neurons were incubated with 10 μg/ml soluble recombinant ICAM-5 D1-4-Fc protein or control mIgG for 72 h. The neurons were then double stained for ICAM-5 (A, red) and MAP-2 (A, blue). Compared with mIgG control protein, sICAM-5 D1-4-Fc protein induced significantly more filopodia from WT neurons, but not from ICAM-5 −/− neurons (B and C). The filopodial length of WT neurons, but not ICAM-5 −/− neurons, also significantly increased in the presence of sICAM-5 D1-4-Fc protein (D). The experiment was repeated three times with similar results. Error bars indicate mean ± SD. *, P

    Journal: The Journal of Cell Biology

    Article Title: Activation of NMDA receptors promotes dendritic spine development through MMP-mediated ICAM-5 cleavage

    doi: 10.1083/jcb.200612097

    Figure Lengend Snippet: sICAM-5 promotes dendritic filopodia elongation. 9-DIV EGFP-transfected WT and ICAM-5 −/− hippocampal neurons were incubated with 10 μg/ml soluble recombinant ICAM-5 D1-4-Fc protein or control mIgG for 72 h. The neurons were then double stained for ICAM-5 (A, red) and MAP-2 (A, blue). Compared with mIgG control protein, sICAM-5 D1-4-Fc protein induced significantly more filopodia from WT neurons, but not from ICAM-5 −/− neurons (B and C). The filopodial length of WT neurons, but not ICAM-5 −/− neurons, also significantly increased in the presence of sICAM-5 D1-4-Fc protein (D). The experiment was repeated three times with similar results. Error bars indicate mean ± SD. *, P

    Article Snippet: Recombinant protein purification Recombinant human ICAM-5 D1-4-Fc and mouse ICAM-5 D1-9-Fc proteins were purified from cell culture supernatants by affinity chromatography with protein A–Sepharose and ÄKTAprime system (GE Healthcare).

    Techniques: Transfection, Incubation, Recombinant, Staining