Journal: Toxicology mechanisms and methods

Article Title: Activation of Protein Kinase C and Protein Kinase D in Human Natural Killer Cells: Effects of Tributyltin, Dibutyltin, and Tetrabromobisphenol A

doi: 10.3109/15376516.2015.1070226

Figure Lengend Snippet: Effects of exposures to TBBPA on the phosphorylation/activation state of PKC and PKD in NK cells: Representative experiment for 10 min exposures to TBBPA. Experiment was repeated using cells from seven different donors.

Article Snippet: NK cells were purified by mixing 30–40 mL of buffy coat with 0.6 mL of RosetteSep human NK cell enrichment antibody cocktail (Stem Cell Technologies, Vancouver, BC, Canada).

Techniques: Activation Assay

Journal: Toxicology mechanisms and methods

Article Title: Activation of Protein Kinase C and Protein Kinase D in Human Natural Killer Cells: Effects of Tributyltin, Dibutyltin, and Tetrabromobisphenol A

doi: 10.3109/15376516.2015.1070226

Figure Lengend Snippet: Effects of incubations with K562 target cells on phosphorylation/activation state of PKC in NK cells: A) levels of phospho-PKC normalized to the control. B) Representative Western blot. Experiments were repeated using cells from five different donors. Asterisks indicate a significant difference as compared with control, p<0.05.

Article Snippet: NK cells were purified by mixing 30–40 mL of buffy coat with 0.6 mL of RosetteSep human NK cell enrichment antibody cocktail (Stem Cell Technologies, Vancouver, BC, Canada).

Techniques: Activation Assay, Western Blot

Journal: Toxicology mechanisms and methods

Article Title: Activation of Protein Kinase C and Protein Kinase D in Human Natural Killer Cells: Effects of Tributyltin, Dibutyltin, and Tetrabromobisphenol A

doi: 10.3109/15376516.2015.1070226

Figure Lengend Snippet: Effects of incubations with K562 target cells on phosphorylation/activation state of PKD in NK cells: A) levels of phospho-PKD normalized to the control. B) Representative Western blot. Experiments were repeated using cells from five different donors. Asterisks indicate a significant difference as compared with control, p<0.05.

Article Snippet: NK cells were purified by mixing 30–40 mL of buffy coat with 0.6 mL of RosetteSep human NK cell enrichment antibody cocktail (Stem Cell Technologies, Vancouver, BC, Canada).

Techniques: Activation Assay, Western Blot

Journal: Toxicology mechanisms and methods

Article Title: Activation of Protein Kinase C and Protein Kinase D in Human Natural Killer Cells: Effects of Tributyltin, Dibutyltin, and Tetrabromobisphenol A

doi: 10.3109/15376516.2015.1070226

Figure Lengend Snippet: Effects of exposures to TBT on the phosphorylation/activation state of PKC in NK cells: A) levels of phospho-PKC normalized to the control for 10 min exposures to TBT. B) Representative experiment for 10 min exposures to TBT. C) Representative experiment for 1 h exposures to TBT. D) Representative experiment for 6 h exposures to TBT. Experiments were repeated using cells from five different donors. Asterisks indicate a significant difference as compared with control, p<0.05.

Article Snippet: NK cells were purified by mixing 30–40 mL of buffy coat with 0.6 mL of RosetteSep human NK cell enrichment antibody cocktail (Stem Cell Technologies, Vancouver, BC, Canada).

Techniques: Activation Assay

Journal: Toxicology mechanisms and methods

Article Title: Activation of Protein Kinase C and Protein Kinase D in Human Natural Killer Cells: Effects of Tributyltin, Dibutyltin, and Tetrabromobisphenol A

doi: 10.3109/15376516.2015.1070226

Figure Lengend Snippet: Effects of exposures to TBT on the phosphorylation/activation state of PKD in NK cells: A) levels of phospho-PKD normalized to the control for 10 min exposures to TBT. B) Representative experiment for 10 min exposures to TBT. C) Representative experiment for 1 h exposures to TBT. D) Representative experiment for 6 h exposures to TBT. Experiments were repeated using cells from five different donors. Asterisks indicate a significant difference as compared with control, p<0.05.

Article Snippet: NK cells were purified by mixing 30–40 mL of buffy coat with 0.6 mL of RosetteSep human NK cell enrichment antibody cocktail (Stem Cell Technologies, Vancouver, BC, Canada).

Techniques: Activation Assay

Journal: Toxicology mechanisms and methods

Article Title: Activation of Protein Kinase C and Protein Kinase D in Human Natural Killer Cells: Effects of Tributyltin, Dibutyltin, and Tetrabromobisphenol A

doi: 10.3109/15376516.2015.1070226

Figure Lengend Snippet: Effects of exposures to DBT on the phosphorylation/activation state of PKC in NK cells: A) levels of phospho-PKC normalized to the control for 10 min exposures to DBT B) Representative experiment for 10 min exposures to DBT. C) levels of phospho-PKC normalized to the control for the 1 h exposures to DBT. D) Representative experiment for 1 h exposures to DBT. E) Representative experiment for 6 h exposures to DBT. Experiments were repeated using cells from six different donors for 10 min and 1h and form three different donors for 6 h. Asterisks indicate a significant difference as compared with control, p<0.05.

Article Snippet: NK cells were purified by mixing 30–40 mL of buffy coat with 0.6 mL of RosetteSep human NK cell enrichment antibody cocktail (Stem Cell Technologies, Vancouver, BC, Canada).

Techniques: Activation Assay

Journal: Toxicology mechanisms and methods

Article Title: Activation of Protein Kinase C and Protein Kinase D in Human Natural Killer Cells: Effects of Tributyltin, Dibutyltin, and Tetrabromobisphenol A

doi: 10.3109/15376516.2015.1070226

Figure Lengend Snippet: Effects of exposures to DBT on the phosphorylation/activation state of PKD in NK cells: A) levels of phospho-PKD normalized to the control for 10 min exposures to DBT. B) Representative experiment for 10 min exposures to DBT. C) Representative experiment for 1 h exposures to DBT. D) Representative experiment for 6 h exposures to DBT. Experiments were repeated using cells from five different donors (10 min), six different donors (1 h) and three different donors (6 h). Asterisks indicate a significant difference as compared with control, p<0.05.

Article Snippet: NK cells were purified by mixing 30–40 mL of buffy coat with 0.6 mL of RosetteSep human NK cell enrichment antibody cocktail (Stem Cell Technologies, Vancouver, BC, Canada).

Techniques: Activation Assay

Journal: Particle and Fibre Toxicology

Article Title: Ultrafine carbon particles down-regulate CYP1B1 expression in human monocytes

doi: 10.1186/1743-8977-6-27

Figure Lengend Snippet: A) MDM were generated from CD14++ monocytes purified from PBMC by MACS separation followed by 5-day incubation with M-CSF (100 ng/ml) . Cells remained untreated (none) or were stimulated with LPS (10 ng/ml) or with particle-mix of ultrafine P90 and fine TiO 2 (each with 32 μg/ml) for 3 h (n = 5 patients with COPD, n = 8 healthy controls, mean ± S.D.; * p < 0.05 compared to untreated controls). B) Effect of particles on CYP1B1 expression in sputum macrophages of healthy non-smokers, healthy smokers and patients with COPD. Sputum macrophages were purified using RosetteSep to deplete unwanted leukocytes. Cells remained untreated (none) or were stimulated with LPS (10 ng/ml) or with particle-mix of ultrafine P90 and fine TiO 2 (each with 32 μg/ml) for 3 h (n = 5 non-smokers, n = 4 smokers, n = 7 patients with COPD, mean ± S.D.; * p < 0.05). C) Cells remained untreated (none) or were stimulated with LPS (10 ng/ml), with particle mix of ultrafine P90 and fine TiO 2 (each with 32 μg/ml) or with each particle separately for 22 h (each with 32 μg/ml) (A549 n = 3, Calu-3 n = 4 experiments from different cell passages, mean ± S.D.; * p < 0.05 compared to untreated control). D) Effect of ultrafine P90 on CYP1B1 mRNA expressin in primary bronchial epithelial cells. Cells were obtained by bronchial brush biopsy. Cells remained untreated (none) or were stimulated with ultrafine P90 (32 μg/ml) for 3 h (n = 7 incubations from different donors, mean ± S.D.; * p < 0.05 compared to untreated control).

Article Snippet: 10 μl packed erythrocytes and 50 μl RosetteSep Human Monocyte Enrichment Cocktail (15068, CellSystems, St. Katharinen, Germany) were added, which crosslinks unwanted cells to multiple red blood cells, forming immunorosettes.

Techniques: Generated, Purification, Incubation, Expressing, Control