prism 2.01 for windows 95 Search Results


cd14 microbeads  (Miltenyi Biotec)
98
Miltenyi Biotec cd14 microbeads
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Cd14 Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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laser l  (Sanyo)
86
Sanyo laser l
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Laser L, supplied by Sanyo, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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phenanthrenequinone  (Chem Impex International)
95
Chem Impex International phenanthrenequinone
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Phenanthrenequinone, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dexmedetomidine  (Croda International Plc)
93
Croda International Plc dexmedetomidine
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Dexmedetomidine, supplied by Croda International Plc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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laser beam l  (Sanyo)
86
Sanyo laser beam l
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Laser Beam L, supplied by Sanyo, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/prism+2%2E01+for+windows+95/beam+l+laser/arxiv__1511__04263-55-2-5
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hf dialyzers  (CH Instruments)
90
CH Instruments hf dialyzers
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Hf Dialyzers, supplied by CH Instruments, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rat igg2a isotype control  (Becton Dickinson)
90
Becton Dickinson rat igg2a isotype control
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Rat Igg2a Isotype Control, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/prism+2%2E01+for+windows+95/isotype+control+antibodies/10__1128_slash_iai__00910___17-92-25-31
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refractometer dr 201-95  (KRUSS GmbH)
90
KRUSS GmbH refractometer dr 201-95
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Refractometer Dr 201 95, supplied by KRUSS GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/prism+2%2E01+for+windows+95/digital+refractometer+dr201+95/pm37507995-62-19-22
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thunderbird probe qpcr mix  (Toyobo)
99
Toyobo thunderbird probe qpcr mix
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Thunderbird Probe Qpcr Mix, supplied by Toyobo, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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biotage chromatography  (BIOTAGE)
90
BIOTAGE biotage chromatography
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Biotage Chromatography, supplied by BIOTAGE, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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kod plus dna polymerase  (Toyobo)
96
Toyobo kod plus dna polymerase
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Kod Plus Dna Polymerase, supplied by Toyobo, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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solid phase resins  (Chem Impex International)
95
Chem Impex International solid phase resins
Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated <t>anti-CD14</t> and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.
Solid Phase Resins, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated anti-CD14 and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.

Journal: Arthritis & rheumatology (Hoboken, N.J.)

Article Title: Dysregulated osteoclastogenesis is related to natural killer T cell dysfunction in rheumatoid arthritis.

doi: 10.1002/art.39244

Figure Lengend Snippet: Figure 5. Decreased phosphorylation of STAT-1 by a-galactosylceramide (aGalCer) in patients with rheumatoid arthritis (RA). A, Western blot analysis of phospho–STAT-1 expression. Monocytes were isolated from healthy control (HC) and RA patient peripheral blood mononuclear cells (PBMCs) by magnetic-activated cell sorting. PBMCs (6 3 105 cells/well) and monocytes (1 3 106 cells/well) were seeded in the upper and lower compartments, respectively, of a Transwell system with 0.4-mm pores. PBMCs in the upper compartment were incubated for 4 hours in the pres- ence of aGalCer (100 ng/ml). After stimulation, monocytes were harvested and Western blotted (top). Levels of phospho–STAT-1 were quanti- fied using an LAS3000 luminescent image analyzer (bottom). B, Flow cytometric analysis of phospho–STAT-1 expression. Freshly isolated PBMCs (1 3 106 cells/well) were incubated in the presence of aGalCer (100 ng/ml) for 24 hours, interferon-g (IFNg; 20 ng/ml) for 15 minutes, or 0.1% DMSO as control and then stained with fluorescein isothiocyanate–conjugated anti-CD14 and phycoerythrin-conjugated anti–phospho– STAT-1 monoclonal antibodies (mAb). Phospho–STAT-1 expression in the monocyte population was determined by intracellular flow cytometry. Black lines, gray lines, and dotted lines represent monocyte phospho–STAT-1 expression in the presence of IFNg, aGalCer, and 0.1% DMSO, respectively. Shaded regions indicate isotype-matched control mAb. Results are representative of 3 independent experiments.

Article Snippet: Monocytes were isolated from PBMCs at purities of .95% using CD14 MicroBeads according to the instructions of the manufacturer (Miltenyi Biotec).

Techniques: Phospho-proteomics, Western Blot, Expressing, Isolation, Control, FACS, Incubation, Staining, Bioprocessing, Flow Cytometry