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PanReac AppliChem dnase i
d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus <t>DNase</t> I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
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Images

1) Product Images from "d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression"

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00975

d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
Figure Legend Snippet: d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

Techniques Used:

PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P
Figure Legend Snippet: PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

Techniques Used: Activation Assay, Immunofluorescence

2) Product Images from "d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression"

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00975

d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
Figure Legend Snippet: d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

Techniques Used:

PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P
Figure Legend Snippet: PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

Techniques Used: Activation Assay, Immunofluorescence

3) Product Images from "d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression"

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00975

d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
Figure Legend Snippet: d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

Techniques Used:

PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P
Figure Legend Snippet: PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

Techniques Used: Activation Assay, Immunofluorescence

4) Product Images from "d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression"

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00975

d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
Figure Legend Snippet: d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

Techniques Used:

PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P
Figure Legend Snippet: PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

Techniques Used: Activation Assay, Immunofluorescence

5) Product Images from "d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression"

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00975

d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
Figure Legend Snippet: d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

Techniques Used:

PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P
Figure Legend Snippet: PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

Techniques Used: Activation Assay, Immunofluorescence

6) Product Images from "d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression"

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00975

d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
Figure Legend Snippet: d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

Techniques Used:

PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P
Figure Legend Snippet: PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

Techniques Used: Activation Assay, Immunofluorescence

7) Product Images from "d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression"

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00975

d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
Figure Legend Snippet: d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

Techniques Used:

PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P
Figure Legend Snippet: PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

Techniques Used: Activation Assay, Immunofluorescence

Related Articles

Nucleic Acid Electrophoresis:

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Article Snippet: For cell lysis, cells were resuspended in lysis buffer [B‐PER reagent in phosphate buffer (Thermo Scientific) supplemented with 500 mM NaCl, 20 mM imidazole, 4 mM phenylmethanesulfonyl fluoride, 1 mM MgCl2 , 2 mg/ml lysozyme, and 0.2 mg/ml DNase I (PanReac AppliChem), volume: 10% of culture volume]. .. The purity of all tested proteins was higher than 90%, as assessed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE; ), in agreement with the study in which these overexpression strains had first been described.

Centrifugation:

Article Title: Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
Article Snippet: Cells were harvested by centrifugation (5,000 g and 4°C for 15 min) and flash‐frozen in liquid nitrogen. .. For cell lysis, cells were resuspended in lysis buffer [B‐PER reagent in phosphate buffer (Thermo Scientific) supplemented with 500 mM NaCl, 20 mM imidazole, 4 mM phenylmethanesulfonyl fluoride, 1 mM MgCl2 , 2 mg/ml lysozyme, and 0.2 mg/ml DNase I (PanReac AppliChem), volume: 10% of culture volume].

Article Title: Recognition motif and mechanism of ripening inhibitory peptides in plant hormone receptor ETR1
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Flow Cytometry:

Article Title: Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
Article Snippet: For cell lysis, cells were resuspended in lysis buffer [B‐PER reagent in phosphate buffer (Thermo Scientific) supplemented with 500 mM NaCl, 20 mM imidazole, 4 mM phenylmethanesulfonyl fluoride, 1 mM MgCl2 , 2 mg/ml lysozyme, and 0.2 mg/ml DNase I (PanReac AppliChem), volume: 10% of culture volume]. .. His‐tagged proteins were purified from cell extracts using His GraviTrap TALON gravity flow columns (GE Healthcare), and the elution buffer was replaced by the respective assay buffer [20 mM potassium phosphate buffer (pH 7.5), 100 mM KCl, 10 mM NaCl, and 5 mM MgCl2 ] using ZEBA™ spin desalting columns with 7 kDa cutoff (Thermo Scientific).

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression
Article Snippet: .. Indeed, addition of Cl-amidine and DNase I each significantly reduced bovine neutrophil adhesion onto vascular endothelial sheets under flow conditions. .. Neither Ar-c155858 nor DNase I affected neutrophil adhesion after TNFα stimulation, demonstrating specificity for the d (−) lactic acid-induced pathway.

Blocking Assay:

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression
Article Snippet: Next, cells were stimulated with 5 mM of d (−) lactic acid with 90 U DNase I (PanReac AppliChem, Darmstadt, Germany) or with vehicle (0.01% ethanol) for 30 min at 37°C. .. The cells were treated with blocking buffer (4% of non-fat milk, 1% BSA, and 0.05% Tween-20 in PBS) for 2 h at RT and incubated with either anti-CD11b antibody (#MCA1425, ABD Serotec, Raleigh, NC, USA), or anti-H4 citrullinated 3 antibody (#07-596, Merck Millipore, Darmstadt, Germany) overnight at 4°C.

Purification:

Article Title: Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
Article Snippet: Paragraph title: Protein expression and purification ... For cell lysis, cells were resuspended in lysis buffer [B‐PER reagent in phosphate buffer (Thermo Scientific) supplemented with 500 mM NaCl, 20 mM imidazole, 4 mM phenylmethanesulfonyl fluoride, 1 mM MgCl2 , 2 mg/ml lysozyme, and 0.2 mg/ml DNase I (PanReac AppliChem), volume: 10% of culture volume].

Article Title: Recognition motif and mechanism of ripening inhibitory peptides in plant hormone receptor ETR1
Article Snippet: Paragraph title: Expression and purification of AtETR1, its C-terminally truncated constructs and AtETR11–307 mutants ... Cell pellets thawed on ice were resuspended by vortexing in ice-cold lysis buffer 1 [pH 8.0, 140 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 1.8 mM KH2 PO4 , 100 g L−1 glycerol, 20 mg L−1 phenylmethylsulfonyl fluoride (PMSF) and 10 mg L−1 DNase I (PanReac AppliChem); 5 mL lysis buffer per 1 g cells] and broken with Constants Cell Disruption System (Constant Systems) at 2.4 kbar and 5 °C.

Incubation:

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression
Article Snippet: Visualization of NET and Detection of CD11b and Citrunillated 3 Histone H4 in NET-Like Structures A total of 1 × 106 neutrophils were incubated with 1 µM Ar-c155858, 200 µM of Cl-amidine or vehicle (0.01% dimethyl sulfoxide) for 60 min at 37°C. .. Next, cells were stimulated with 5 mM of d (−) lactic acid with 90 U DNase I (PanReac AppliChem, Darmstadt, Germany) or with vehicle (0.01% ethanol) for 30 min at 37°C.

Article Title: Recognition motif and mechanism of ripening inhibitory peptides in plant hormone receptor ETR1
Article Snippet: After incubation for additional 5 h, cells were spun down at 7,500 g for 15 min at 4 °C, flash-frozen in liquid nitrogen and stored at −20 °C. .. Cell pellets thawed on ice were resuspended by vortexing in ice-cold lysis buffer 1 [pH 8.0, 140 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 1.8 mM KH2 PO4 , 100 g L−1 glycerol, 20 mg L−1 phenylmethylsulfonyl fluoride (PMSF) and 10 mg L−1 DNase I (PanReac AppliChem); 5 mL lysis buffer per 1 g cells] and broken with Constants Cell Disruption System (Constant Systems) at 2.4 kbar and 5 °C.

other:

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression
Article Snippet: Similarly, treatment with a PAD4 inhibitor (200 µM, Cl-amidine) and DNase I (90 U) reduced neutrophil NET formation, suggesting that d (−) lactic acid-induced NET formation was PAD4- and MCT1-dependant (Figures A,B).

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression
Article Snippet: Briefly, BUVECs were treated with 5 mM of d (−) lactic acid, vehicle (pH buffer), or with supernatants from neutrophils exposed to 5 mM of d (−) lactic acid, DNase I, or 0.01% vehicle (ethanol) for 30 min at 37°C.

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression
Article Snippet: Neither Ar-c155858 nor DNase I affected neutrophil adhesion after TNFα stimulation, demonstrating specificity for the d (−) lactic acid-induced pathway.

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression
Article Snippet: Moreover, DNase I and Ar-c155858 treatments specifically inhibited d (−) lactic acid-induced neutrophil adhesions.

Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression
Article Snippet: Co-stimulation with DNase I and d (−) lactic acid treatments also significantly decreased neutrophil adhesions onto bovine endothelial sheets.

Construct:

Article Title: Recognition motif and mechanism of ripening inhibitory peptides in plant hormone receptor ETR1
Article Snippet: Paragraph title: Expression and purification of AtETR1, its C-terminally truncated constructs and AtETR11–307 mutants ... Cell pellets thawed on ice were resuspended by vortexing in ice-cold lysis buffer 1 [pH 8.0, 140 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 1.8 mM KH2 PO4 , 100 g L−1 glycerol, 20 mg L−1 phenylmethylsulfonyl fluoride (PMSF) and 10 mg L−1 DNase I (PanReac AppliChem); 5 mL lysis buffer per 1 g cells] and broken with Constants Cell Disruption System (Constant Systems) at 2.4 kbar and 5 °C.

Expressing:

Article Title: Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
Article Snippet: Paragraph title: Protein expression and purification ... For cell lysis, cells were resuspended in lysis buffer [B‐PER reagent in phosphate buffer (Thermo Scientific) supplemented with 500 mM NaCl, 20 mM imidazole, 4 mM phenylmethanesulfonyl fluoride, 1 mM MgCl2 , 2 mg/ml lysozyme, and 0.2 mg/ml DNase I (PanReac AppliChem), volume: 10% of culture volume].

Article Title: Recognition motif and mechanism of ripening inhibitory peptides in plant hormone receptor ETR1
Article Snippet: Paragraph title: Expression and purification of AtETR1, its C-terminally truncated constructs and AtETR11–307 mutants ... Cell pellets thawed on ice were resuspended by vortexing in ice-cold lysis buffer 1 [pH 8.0, 140 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 1.8 mM KH2 PO4 , 100 g L−1 glycerol, 20 mg L−1 phenylmethylsulfonyl fluoride (PMSF) and 10 mg L−1 DNase I (PanReac AppliChem); 5 mL lysis buffer per 1 g cells] and broken with Constants Cell Disruption System (Constant Systems) at 2.4 kbar and 5 °C.

Lysis:

Article Title: Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
Article Snippet: .. For cell lysis, cells were resuspended in lysis buffer [B‐PER reagent in phosphate buffer (Thermo Scientific) supplemented with 500 mM NaCl, 20 mM imidazole, 4 mM phenylmethanesulfonyl fluoride, 1 mM MgCl2 , 2 mg/ml lysozyme, and 0.2 mg/ml DNase I (PanReac AppliChem), volume: 10% of culture volume]. .. The suspension was shaken at room temperature for 10 min, and cell extracts were separated from cell debris by centrifugation (20,000 g and 4°C for 30 min).

Article Title: Recognition motif and mechanism of ripening inhibitory peptides in plant hormone receptor ETR1
Article Snippet: .. Cell pellets thawed on ice were resuspended by vortexing in ice-cold lysis buffer 1 [pH 8.0, 140 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 1.8 mM KH2 PO4 , 100 g L−1 glycerol, 20 mg L−1 phenylmethylsulfonyl fluoride (PMSF) and 10 mg L−1 DNase I (PanReac AppliChem); 5 mL lysis buffer per 1 g cells] and broken with Constants Cell Disruption System (Constant Systems) at 2.4 kbar and 5 °C. .. Cell debris and inclusion bodies were removed by centrifugation at 14,000 g for 30 min.

Over Expression:

Article Title: Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
Article Snippet: Protein expression and purification LB shake flask cultures (200–600 ml, 5 g/l yeast extract, 10 g/l tryptone, 10 g/l NaCl) supplemented with 100 μg/ml chloramphenicol were inoculated in a 1:100 ratio with LB precultures of the overexpression strains (Kitagawa et al , ), and expression was induced with 0.1 mM isopropyl β‐D‐1‐thiogalactopyranoside. .. For cell lysis, cells were resuspended in lysis buffer [B‐PER reagent in phosphate buffer (Thermo Scientific) supplemented with 500 mM NaCl, 20 mM imidazole, 4 mM phenylmethanesulfonyl fluoride, 1 mM MgCl2 , 2 mg/ml lysozyme, and 0.2 mg/ml DNase I (PanReac AppliChem), volume: 10% of culture volume].

SDS Page:

Article Title: Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
Article Snippet: For cell lysis, cells were resuspended in lysis buffer [B‐PER reagent in phosphate buffer (Thermo Scientific) supplemented with 500 mM NaCl, 20 mM imidazole, 4 mM phenylmethanesulfonyl fluoride, 1 mM MgCl2 , 2 mg/ml lysozyme, and 0.2 mg/ml DNase I (PanReac AppliChem), volume: 10% of culture volume]. .. The purity of all tested proteins was higher than 90%, as assessed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE; ), in agreement with the study in which these overexpression strains had first been described.

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    PanReac AppliChem dnase i
    d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus <t>DNase</t> I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P
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    d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

    Journal: Frontiers in Immunology

    Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

    doi: 10.3389/fimmu.2017.00975

    Figure Lengend Snippet: d (−) lactic acid induced the release of cell-free DNA from neutrophils that activated bovine umbilical vein endothelial cell (BUVEC) sheets by ICAM display. (A) Supernatants collected from neutrophils treated with 5 mM d (−) lactic acid plus or minus DNase I were perfused onto BUVEC for 10 min. Next, fresh, untreated neutrophils were perfused on endothelial monolayers, n = 5, ** P

    Article Snippet: Briefly, BUVECs were treated with 5 mM of d (−) lactic acid, vehicle (pH buffer), or with supernatants from neutrophils exposed to 5 mM of d (−) lactic acid, DNase I, or 0.01% vehicle (ethanol) for 30 min at 37°C.

    Techniques:

    PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

    Journal: Frontiers in Immunology

    Article Title: d(−) Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

    doi: 10.3389/fimmu.2017.00975

    Figure Lengend Snippet: PAD4 activation regulates d (−) lactic acid-induced neutrophil extracellular trap (NET) formation and subsequent adhesion. (A) Immunofluorescence of bovine neutrophils pre-treated with monocarboxylate transporter 1 inhibitor (Ar-c155858), PAD4 inhibitor (Cl-amidine), or vehicle (0.01% DMSO) for 60 min, followed by stimulation with either 5 mM d (−) lactic acid or vehicle concomitant with DNase I (90 U) or controls for 30 min. Neutrophils were probed with anti-H 4 citrullinated 3/alexa 405, CD11b/alexa 635, and PicoGreen as a probe for DNA; representative images from three independent experiments, scale bar: 50 µm. (B) Fold of control [(number of NETs/number of neutrophils) × 100] from neutrophils treated with 1 µM Ar-c155858, 200 µM Cl-amidine or vehicle for 1 h and stimulated with 5 mM of d (−) lactic acid or vehicle (control) together or not with DNAse I (90 U) for 30 min, n = 3. *** P

    Article Snippet: Briefly, BUVECs were treated with 5 mM of d (−) lactic acid, vehicle (pH buffer), or with supernatants from neutrophils exposed to 5 mM of d (−) lactic acid, DNase I, or 0.01% vehicle (ethanol) for 30 min at 37°C.

    Techniques: Activation Assay, Immunofluorescence