Expressing:Article Title: Non-ATP/catalytic site p38 mitogen activated protein kinase inhibitors
Article Snippet: Cytotoxicity of HMVECL exposed to 10-100 μM of each compound is analyzed by MTS assay (Promega), LDH release (Promega), and immunoblotting for activated caspase-3 (Cell Signaling). .. Macrophage cytokine expression: The capacity of test compounds to block LPS-induced cytokine expression is assessed in PMA-differentiated THP1 cells using qRT-PCR and Luminex-based immunoassays (UMB Cytokine Core Lab). .. THP1 cells differentiated with 5 ng/ml PMA for 24 h, are treated with 1-100 μM test compound, 10 μM SB203580, or DMSO for 1 h, then with 100 ng/ml ultrapure E. coli 0111:B4 LPS (InvivoGen) for 3 h (qRT-PCR; Real Time Primers) or 24 h (supernatants for immunoassays).
Blocking Assay:Article Title: Non-ATP/catalytic site p38 mitogen activated protein kinase inhibitors
Article Snippet: Cytotoxicity of HMVECL exposed to 10-100 μM of each compound is analyzed by MTS assay (Promega), LDH release (Promega), and immunoblotting for activated caspase-3 (Cell Signaling). .. Macrophage cytokine expression: The capacity of test compounds to block LPS-induced cytokine expression is assessed in PMA-differentiated THP1 cells using qRT-PCR and Luminex-based immunoassays (UMB Cytokine Core Lab). .. THP1 cells differentiated with 5 ng/ml PMA for 24 h, are treated with 1-100 μM test compound, 10 μM SB203580, or DMSO for 1 h, then with 100 ng/ml ultrapure E. coli 0111:B4 LPS (InvivoGen) for 3 h (qRT-PCR; Real Time Primers) or 24 h (supernatants for immunoassays).
Quantitative RT-PCR:Article Title: Non-ATP/catalytic site p38 mitogen activated protein kinase inhibitors
Article Snippet: Cytotoxicity of HMVECL exposed to 10-100 μM of each compound is analyzed by MTS assay (Promega), LDH release (Promega), and immunoblotting for activated caspase-3 (Cell Signaling). .. Macrophage cytokine expression: The capacity of test compounds to block LPS-induced cytokine expression is assessed in PMA-differentiated THP1 cells using qRT-PCR and Luminex-based immunoassays (UMB Cytokine Core Lab). .. THP1 cells differentiated with 5 ng/ml PMA for 24 h, are treated with 1-100 μM test compound, 10 μM SB203580, or DMSO for 1 h, then with 100 ng/ml ultrapure E. coli 0111:B4 LPS (InvivoGen) for 3 h (qRT-PCR; Real Time Primers) or 24 h (supernatants for immunoassays).
Concentration Assay:Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions.
Article Snippet: .. THP1 cells, at a concentration of 1 × 105 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4, Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. After incubation, 20 μL of culture supernatant was transferred to a new 96-well plate and mixed with 180 μL of QUANTI-Blue Solution (InvivoGen, France).
Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions
Article Snippet: .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules).
Incubation:Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions.
Article Snippet: .. THP1 cells, at a concentration of 1 × 105 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4, Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. After incubation, 20 μL of culture supernatant was transferred to a new 96-well plate and mixed with 180 μL of QUANTI-Blue Solution (InvivoGen, France).
Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions
Article Snippet: .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules).
Positive Control:Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions.
Article Snippet: .. THP1 cells, at a concentration of 1 × 105 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4, Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. After incubation, 20 μL of culture supernatant was transferred to a new 96-well plate and mixed with 180 μL of QUANTI-Blue Solution (InvivoGen, France).
Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions
Article Snippet: .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules).
Control:Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions.
Article Snippet: .. THP1 cells, at a concentration of 1 × 105 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4, Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. After incubation, 20 μL of culture supernatant was transferred to a new 96-well plate and mixed with 180 μL of QUANTI-Blue Solution (InvivoGen, France).
Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions
Article Snippet: .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules).
Capsules:Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions.
Article Snippet: .. THP1 cells, at a concentration of 1 × 105 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4, Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. After incubation, 20 μL of culture supernatant was transferred to a new 96-well plate and mixed with 180 μL of QUANTI-Blue Solution (InvivoGen, France).
Article Title: Differential stress responses of immunoisolated human islets embedded in pancreatic extracellular matrix under static and free-fall dynamic conditions
Article Snippet: .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules). .. THP1 cells, at a concentration of 1 × 10 5 cells per well in a 96-well plate, were incubated for 24 h with either KRH buffer, 10 ng/mL lipopolysaccharides (LPS from E. coli O111:B4 , Invivogen, France, cat#tlrl-3pelps) as a positive control, or varying amounts of empty microcapsules (5, 10, or 20 control alginate capsules or alginate with ECM capsules).
Cell Culture:Article Title: NLRP3 inhibition by VTX3232 tempers inflammation resulting in reduced body weight, hyperglycemia, and hepatic steatosis in obese male mice
Article Snippet: Chromatographic separation was achieved on a Waters ACQUITY UPLC CSH C18 column (1.7 μm, 2.1 × 50 mm) at room temperature using a gradient mobile phase system of water and acetonitrile (both with 0.1% formic acid) at 0.6 mL/min flow rate over 2.5 min. Mass spectrometry detection employed positive ionization mode with MRM transitions of 271.10 → 91.10 for tolbutamide (internal standard) and 382.20 → 255.20 for VTX3232, with optimized source parameters and a 2.0 μL injection volume. .. THP1 cells (Invivogen, thpd-nfis; purchased as mycoplasma-free THP1 cells and regularly tested for mycoplasma) were cultured in RPMI-1640 GlutaMAX (Gibco, 72400047) supplemented with 10 % FBS (Gibco, A5256801), 1x penicillin/streptomycin (Gibco, 15140122), 100 μg/mL Normocin (InvivoGen, ant-nr-1), 10 μg/mL Blasticidin S (Gibco, A11139-03), and 100 μg/mL Zeocin (InvivoGen, ant-zn-05). ..
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