Cell Culture:Article Title: PARP Traps Rescue the Pro-Inflammatory Response of Human Macrophages in the In Vitro Model of LPS-Induced Tolerance
Article Snippet: THP1 cells and human monocytes were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA) and from healthy donors from the blood bank in Lodz, respectively. .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France); cell culture media were from Biowest (CytoGen, Zgierz, Poland); granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK); BLUeye prestained protein ladder (#94964), oligonucleotides for real-time PCR were from Sigma Aldrich (Poznan, Poland); 5x HOT FIREPol EvaGreen qPCR Mix Plus (no ROX) (CytoGen, Zgierz Poland), ChiP grade antibodies anti-histone H3 (#4620), anti-PARP1 (#9532) and normal rabbit IgG (#2729) were from Cell Signaling Technology (LabJOT, Warsaw, Poland); TRI Reagent, anti-rabbit IgG (A0545) (whole molecule)–peroxidase antibody produced in goat were from Sigma Aldrich (Poznan, Poland); Lipofectamine RNAiMAX, DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, SuperSignalTM West Pico Chemiluminescent Substrate, OptiMem were from Thermo Fisher Scientific (Warsaw, Poland). .. Lipopolysaccharide (LPS), PARG inhibitor—ADP-HPD (Dihydrate, Ammonium Salt–Calbiochem) were from Sigma Aldrich (Poznan, Poland), Olaparib, Niraparib (MK-4827) and Veliparib (ABT-888), MG-132 and iPARG were from Cayman Chemical Biokom, (Warsaw, Poland).
Article Title: LSD1 Facilitates Pro-Inflammatory Polarization of Macrophages by Repressing Catalase
Article Snippet: .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France), cell culture media were from Biowest (CytoGen, Zgierz, Poland), granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK), anti-rabbit IgG (whole molecule) (A0545) and anti-mouse IgG (whole molecule) (A4416) peroxidase-labeled antibodies produced in goat, BLUeye prestained protein marker (#94964), SIGMAFASTTM Protease Inhibitor Tablets (PIC), sodium butyrate (iHDAC), C646 (iEP300), ML385 (iNRF2) and oligonucleotides for real-time PCR were from Sigma-Aldrich (Poznan, Poland). .. DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, Click-iTTM Nascent RNA Capture Kit, SuperSignalTM West Pico Chemiluminescent Substrate, TRI ReagentTM, PowerUpTM SYBR ® Green Master Mix, and anty-HDAC1 (PA1-860) were from Thermofisher Scientific (Thermofisher Scientific, Warsaw, Poland).
Article Title: EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages.
Article Snippet: Accepted Manuscript EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages Julita Pietrzak, Tomasz Płoszaj, Łukasz Pułaski, Agnieszka Robaszkiewicz PII: S1874-9399(18)30342-0 DOI: https://doi.org/10.1016/j.bbagrm.2018.10.019 Reference: BBAGRM 94334 To appear in: BBA - Gene Regulatory Mechanisms Received date: 20 August 2018 Revised date: 17 October 2018 Accepted date: 31 October 2018 Please cite this article as: Julita Pietrzak, Tomasz Płoszaj, Łukasz Pułaski, Agnieszka Robaszkiewicz , EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages.. Bbagrm (2018), https://doi.org/10.1016/j.bbagrm.2018.10.019 This is a PDF file of an unedited manuscript that has been accepted for publication.. As a service to our customers we are providing this early version of the manuscript.
Real-time Polymerase Chain Reaction:Article Title: PARP Traps Rescue the Pro-Inflammatory Response of Human Macrophages in the In Vitro Model of LPS-Induced Tolerance
Article Snippet: THP1 cells and human monocytes were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA) and from healthy donors from the blood bank in Lodz, respectively. .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France); cell culture media were from Biowest (CytoGen, Zgierz, Poland); granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK); BLUeye prestained protein ladder (#94964), oligonucleotides for real-time PCR were from Sigma Aldrich (Poznan, Poland); 5x HOT FIREPol EvaGreen qPCR Mix Plus (no ROX) (CytoGen, Zgierz Poland), ChiP grade antibodies anti-histone H3 (#4620), anti-PARP1 (#9532) and normal rabbit IgG (#2729) were from Cell Signaling Technology (LabJOT, Warsaw, Poland); TRI Reagent, anti-rabbit IgG (A0545) (whole molecule)–peroxidase antibody produced in goat were from Sigma Aldrich (Poznan, Poland); Lipofectamine RNAiMAX, DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, SuperSignalTM West Pico Chemiluminescent Substrate, OptiMem were from Thermo Fisher Scientific (Warsaw, Poland). .. Lipopolysaccharide (LPS), PARG inhibitor—ADP-HPD (Dihydrate, Ammonium Salt–Calbiochem) were from Sigma Aldrich (Poznan, Poland), Olaparib, Niraparib (MK-4827) and Veliparib (ABT-888), MG-132 and iPARG were from Cayman Chemical Biokom, (Warsaw, Poland).
Article Title: LSD1 Facilitates Pro-Inflammatory Polarization of Macrophages by Repressing Catalase
Article Snippet: .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France), cell culture media were from Biowest (CytoGen, Zgierz, Poland), granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK), anti-rabbit IgG (whole molecule) (A0545) and anti-mouse IgG (whole molecule) (A4416) peroxidase-labeled antibodies produced in goat, BLUeye prestained protein marker (#94964), SIGMAFASTTM Protease Inhibitor Tablets (PIC), sodium butyrate (iHDAC), C646 (iEP300), ML385 (iNRF2) and oligonucleotides for real-time PCR were from Sigma-Aldrich (Poznan, Poland). .. DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, Click-iTTM Nascent RNA Capture Kit, SuperSignalTM West Pico Chemiluminescent Substrate, TRI ReagentTM, PowerUpTM SYBR ® Green Master Mix, and anty-HDAC1 (PA1-860) were from Thermofisher Scientific (Thermofisher Scientific, Warsaw, Poland).
Article Title: EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages.
Article Snippet: Accepted Manuscript EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages Julita Pietrzak, Tomasz Płoszaj, Łukasz Pułaski, Agnieszka Robaszkiewicz PII: S1874-9399(18)30342-0 DOI: https://doi.org/10.1016/j.bbagrm.2018.10.019 Reference: BBAGRM 94334 To appear in: BBA - Gene Regulatory Mechanisms Received date: 20 August 2018 Revised date: 17 October 2018 Accepted date: 31 October 2018 Please cite this article as: Julita Pietrzak, Tomasz Płoszaj, Łukasz Pułaski, Agnieszka Robaszkiewicz , EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages.. Bbagrm (2018), https://doi.org/10.1016/j.bbagrm.2018.10.019 This is a PDF file of an unedited manuscript that has been accepted for publication.. As a service to our customers we are providing this early version of the manuscript.
Produced:Article Title: PARP Traps Rescue the Pro-Inflammatory Response of Human Macrophages in the In Vitro Model of LPS-Induced Tolerance
Article Snippet: THP1 cells and human monocytes were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA) and from healthy donors from the blood bank in Lodz, respectively. .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France); cell culture media were from Biowest (CytoGen, Zgierz, Poland); granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK); BLUeye prestained protein ladder (#94964), oligonucleotides for real-time PCR were from Sigma Aldrich (Poznan, Poland); 5x HOT FIREPol EvaGreen qPCR Mix Plus (no ROX) (CytoGen, Zgierz Poland), ChiP grade antibodies anti-histone H3 (#4620), anti-PARP1 (#9532) and normal rabbit IgG (#2729) were from Cell Signaling Technology (LabJOT, Warsaw, Poland); TRI Reagent, anti-rabbit IgG (A0545) (whole molecule)–peroxidase antibody produced in goat were from Sigma Aldrich (Poznan, Poland); Lipofectamine RNAiMAX, DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, SuperSignalTM West Pico Chemiluminescent Substrate, OptiMem were from Thermo Fisher Scientific (Warsaw, Poland). .. Lipopolysaccharide (LPS), PARG inhibitor—ADP-HPD (Dihydrate, Ammonium Salt–Calbiochem) were from Sigma Aldrich (Poznan, Poland), Olaparib, Niraparib (MK-4827) and Veliparib (ABT-888), MG-132 and iPARG were from Cayman Chemical Biokom, (Warsaw, Poland).
Article Title: LSD1 Facilitates Pro-Inflammatory Polarization of Macrophages by Repressing Catalase
Article Snippet: .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France), cell culture media were from Biowest (CytoGen, Zgierz, Poland), granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK), anti-rabbit IgG (whole molecule) (A0545) and anti-mouse IgG (whole molecule) (A4416) peroxidase-labeled antibodies produced in goat, BLUeye prestained protein marker (#94964), SIGMAFASTTM Protease Inhibitor Tablets (PIC), sodium butyrate (iHDAC), C646 (iEP300), ML385 (iNRF2) and oligonucleotides for real-time PCR were from Sigma-Aldrich (Poznan, Poland). .. DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, Click-iTTM Nascent RNA Capture Kit, SuperSignalTM West Pico Chemiluminescent Substrate, TRI ReagentTM, PowerUpTM SYBR ® Green Master Mix, and anty-HDAC1 (PA1-860) were from Thermofisher Scientific (Thermofisher Scientific, Warsaw, Poland).
Article Title: EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages.
Article Snippet: Accepted Manuscript EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages Julita Pietrzak, Tomasz Płoszaj, Łukasz Pułaski, Agnieszka Robaszkiewicz PII: S1874-9399(18)30342-0 DOI: https://doi.org/10.1016/j.bbagrm.2018.10.019 Reference: BBAGRM 94334 To appear in: BBA - Gene Regulatory Mechanisms Received date: 20 August 2018 Revised date: 17 October 2018 Accepted date: 31 October 2018 Please cite this article as: Julita Pietrzak, Tomasz Płoszaj, Łukasz Pułaski, Agnieszka Robaszkiewicz , EP300-HDAC1-SWI/SNF functional unit defines transcription of some DNA repair enzymes during differentiation of human macrophages.. Bbagrm (2018), https://doi.org/10.1016/j.bbagrm.2018.10.019 This is a PDF file of an unedited manuscript that has been accepted for publication.. As a service to our customers we are providing this early version of the manuscript.
Reverse Transcription:Article Title: PARP Traps Rescue the Pro-Inflammatory Response of Human Macrophages in the In Vitro Model of LPS-Induced Tolerance
Article Snippet: THP1 cells and human monocytes were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA) and from healthy donors from the blood bank in Lodz, respectively. .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France); cell culture media were from Biowest (CytoGen, Zgierz, Poland); granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK); BLUeye prestained protein ladder (#94964), oligonucleotides for real-time PCR were from Sigma Aldrich (Poznan, Poland); 5x HOT FIREPol EvaGreen qPCR Mix Plus (no ROX) (CytoGen, Zgierz Poland), ChiP grade antibodies anti-histone H3 (#4620), anti-PARP1 (#9532) and normal rabbit IgG (#2729) were from Cell Signaling Technology (LabJOT, Warsaw, Poland); TRI Reagent, anti-rabbit IgG (A0545) (whole molecule)–peroxidase antibody produced in goat were from Sigma Aldrich (Poznan, Poland); Lipofectamine RNAiMAX, DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, SuperSignalTM West Pico Chemiluminescent Substrate, OptiMem were from Thermo Fisher Scientific (Warsaw, Poland). .. Lipopolysaccharide (LPS), PARG inhibitor—ADP-HPD (Dihydrate, Ammonium Salt–Calbiochem) were from Sigma Aldrich (Poznan, Poland), Olaparib, Niraparib (MK-4827) and Veliparib (ABT-888), MG-132 and iPARG were from Cayman Chemical Biokom, (Warsaw, Poland).
Isolation:Article Title: Engineering the bone metastatic prostate cancer niche through a microphysiological system to report patient-specific treatment response.
Article Snippet: The cell pellet was washed twice with PBS and then cultured in Prostate Epithelial Cell media (PEpiCM, ScienCell). .. For monocytes, peripheral blood from prostate cancer donors was collected and monocytes were isolated using RosetteSepTM Monocyte Enrichment Cocktail (Stemcell) and following the manufacturer protocol. ..
Marker:Article Title: LSD1 Facilitates Pro-Inflammatory Polarization of Macrophages by Repressing Catalase
Article Snippet: .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France), cell culture media were from Biowest (CytoGen, Zgierz, Poland), granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK), anti-rabbit IgG (whole molecule) (A0545) and anti-mouse IgG (whole molecule) (A4416) peroxidase-labeled antibodies produced in goat, BLUeye prestained protein marker (#94964), SIGMAFASTTM Protease Inhibitor Tablets (PIC), sodium butyrate (iHDAC), C646 (iEP300), ML385 (iNRF2) and oligonucleotides for real-time PCR were from Sigma-Aldrich (Poznan, Poland). .. DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, Click-iTTM Nascent RNA Capture Kit, SuperSignalTM West Pico Chemiluminescent Substrate, TRI ReagentTM, PowerUpTM SYBR ® Green Master Mix, and anty-HDAC1 (PA1-860) were from Thermofisher Scientific (Thermofisher Scientific, Warsaw, Poland).
Protease Inhibitor:Article Title: LSD1 Facilitates Pro-Inflammatory Polarization of Macrophages by Repressing Catalase
Article Snippet: .. RosetteSepTM Monocyte Enrichment Cocktail was purchased from STEMCELL Technologies (Grenoble, France), cell culture media were from Biowest (CytoGen, Zgierz, Poland), granulocyte–macrophage colony-stimulating factor (GM-CSF) from PeproTech (London, UK), anti-rabbit IgG (whole molecule) (A0545) and anti-mouse IgG (whole molecule) (A4416) peroxidase-labeled antibodies produced in goat, BLUeye prestained protein marker (#94964), SIGMAFASTTM Protease Inhibitor Tablets (PIC), sodium butyrate (iHDAC), C646 (iEP300), ML385 (iNRF2) and oligonucleotides for real-time PCR were from Sigma-Aldrich (Poznan, Poland). .. DynabeadsTM Protein G, High-Capacity cDNA Reverse Transcription Kit, Click-iTTM Nascent RNA Capture Kit, SuperSignalTM West Pico Chemiluminescent Substrate, TRI ReagentTM, PowerUpTM SYBR ® Green Master Mix, and anty-HDAC1 (PA1-860) were from Thermofisher Scientific (Thermofisher Scientific, Warsaw, Poland).
Gentle:Article Title: Evidence for the presence of air pollution nanoparticles in placental tissue cells.
Article Snippet: Briefly, for erythrocyte extraction, blood was aspirated from the umbilical cord on the day of processing and diluted with 0.6 mL incomplete Roswell Park Memorial Institute (RPMI- 1640, Gibco, Thermo Fisher Scientific, USA, 4% FBS) per 1 mL of blood, before suspension (1:1) on density gradient medium, Ficoll-PaquePlus (GE Healthcare, Sweden), and centrifugation at 1821 rpm (650 xg) for 22 min. Erythrocytes were collected as a pellet. .. For each mL of resuspended placental cells, 30 μL of erythrocytes and 50 μL of RosetteSepTM monocyte enrichment cocktail (STEMCELL Technologies) was added before gentle shaking at room temperature for 20 min. .. The sample was diluted with DPBS/FBS and suspendedon Ficoll-Plaque density gradientmedium(GEHealthcare, Sweden) (1:1).
Purification:Article Title: Activation of caspase-1 is mediated by stimulation of interferon genes and NLR family pyrin domain containing 3 in monocytes of active systemic lupus erythematosus.
Article Snippet: Peripheral blood from HC was collected in BD VacutainerTM Glass ACD Solution A tube (Becton Dickson). .. Monocytes were purified using RosetteSepTM Monocyte Enrichment Cocktail (STEMCELL Technologies) in accordance with the manufacturer’s instruction. ..
In Vitro:Article Title: Single stranded oligonucleotides inhibiting endocytosis
Article Snippet: .. Human In Vitro Derived DCs Monocytes were negatively selected from buffy coats using the RosetteSepTM Monocyte Enrichment Cocktail (StemCell Technologies) and differentiated into DC, as described previously (Sköld, A. E. et al 2012 Blood 120:768-777) at a density of 5×105 cells/mL in RPMI 1640 completed with 10% FCS, 1 mM sodium pyruvate, 10 mM HEPES, 2 mM L-glutamine, and 1% streptomycin and penicillin (all from Invitrogen Life Technologies), with GM-CSF (250 ng/mL; PeproTech) and IL-4 (6.5 ng/mL; R&D Systems) for 6 or 7 days. .. 1×105 MoDCs were exposed to Cy3 labeled ploy I:C (polyI:C was labeled using Mirusbio LabelIT® Nucleic Acid Labeling Kit 3600 according to manufacturer's protocol) and MedON oligonucleotides on in 100 μl complete 10% RPMI media, at 37° C. or +4° C. for 45 minutes.
Derivative Assay:Article Title: Single stranded oligonucleotides inhibiting endocytosis
Article Snippet: .. Human In Vitro Derived DCs Monocytes were negatively selected from buffy coats using the RosetteSepTM Monocyte Enrichment Cocktail (StemCell Technologies) and differentiated into DC, as described previously (Sköld, A. E. et al 2012 Blood 120:768-777) at a density of 5×105 cells/mL in RPMI 1640 completed with 10% FCS, 1 mM sodium pyruvate, 10 mM HEPES, 2 mM L-glutamine, and 1% streptomycin and penicillin (all from Invitrogen Life Technologies), with GM-CSF (250 ng/mL; PeproTech) and IL-4 (6.5 ng/mL; R&D Systems) for 6 or 7 days. .. 1×105 MoDCs were exposed to Cy3 labeled ploy I:C (polyI:C was labeled using Mirusbio LabelIT® Nucleic Acid Labeling Kit 3600 according to manufacturer's protocol) and MedON oligonucleotides on in 100 μl complete 10% RPMI media, at 37° C. or +4° C. for 45 minutes.
Incubation:Article Title: Carbon-Based Nanomaterials Increase Reactivity of Primary Monocytes towards Various Bacteria and Modulate Their Differentiation into Macrophages
Article Snippet: .. Following incubation with 50 μL/mL of RosetteSepTM Monocyte Enrichment Cocktail (STEMCELL Technologies Inc., Vancouver, Canada), gradient density centrifugation (1200× g , 20 min, RT) was performed using Histopaque ® -1077 (Sigma-Aldrich, St. Luis, MO, USA). .. The purity (~85%, ) of isolated monocytes was evaluated by flow cytometry (NaviosTM, Beckman Coulter, Brea, CA, USA).
Centrifugation:Article Title: Carbon-Based Nanomaterials Increase Reactivity of Primary Monocytes towards Various Bacteria and Modulate Their Differentiation into Macrophages
Article Snippet: .. Following incubation with 50 μL/mL of RosetteSepTM Monocyte Enrichment Cocktail (STEMCELL Technologies Inc., Vancouver, Canada), gradient density centrifugation (1200× g , 20 min, RT) was performed using Histopaque ® -1077 (Sigma-Aldrich, St. Luis, MO, USA). .. The purity (~85%, ) of isolated monocytes was evaluated by flow cytometry (NaviosTM, Beckman Coulter, Brea, CA, USA).
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