SDS Page:Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t
Article Snippet: .. Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards, and Triton X-100 (161-0324, 161-0407; BioRad, Hercules, CA); S -BEL (iPLA 2 β inhibitor), PGE 2 (10006801, 14010; Cayman Chemicals, Ann Arbor, MI); acetyl histone H4 (AcH4antibody, normal mouse IgG, protein A beads (06-911, 12-371, 16-125; EMD Millipore, Billerica, MA); Collagenase 2, fetal bovine serum (FBS) and common cell culture reagents (17101-015; Gibco, Carlsbad, CA); Immobilin-P PVDF membrane (IPVH00010; Millipore Corp., Bedford, MA; protease inhibitor cocktail (D4693), alpha-MEM media (P8340), and common reagents (Sigma Chemical Co., St. Louis, MO); and enhanced chemiluminescence reagent, (34095; Thermo-Fisher, Waltham, MA). .. Rat-derived INS-1 β-cells were cultured in Roswell Park Memorial Institute (RPMI, Cat. 11875093, Thermo Fisher Scientific Inc. Waltham, MA ) 1640 medium, containing 11 mM glucose, 10% fetal calf serum, 10 mM HEPES buffer, 2 mM glutamine, 1 mM sodium pyruvate, 50 mM β-mercaptoethanol (BME), and 0.1% (w/v) each of penicillin and streptomycin in cell culture conditions (37°C, 5%CO 2 /95% air).
Article Title: Ca 2+ -independent phospholipase A 2 β-derived PGE 2 contributes to osteogenesis.
Article Snippet: .. Reagents used in our studies were obtained from the following sources (catalog numbers): Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards (161− 0324), and Triton X-100 (161− 0407) from BioRad, Hercules, CA; S-BEL (iPLA2β inhibitor, 10,006,801), R-BEL (iPLA2γ inhibitor, 10,006,800), CAY10502 (cPLA2α inhibitor, 10,008,657), PGE2 (14,010), and PGE2 EIA Kit (514,010) from Cayman Chemicals, Ann Arbor, MI); paraformaldehyde from Electron Microscopy Sciences, Ft. Washington, PA; acetyl histone H3 antibody (06–911), normal mouse IgG (12–371), protein A beads (16–125) from EMD Millipore, Billerica, MA; Immobilon-P PVDF membrane (IPVH00010) from Millipore Corp., Bedford, MA; Runx2 antibody (D130− 3) from MBL International, Wobrurn, MA); arachidonic acid (ICN 19,462,510) from MP Bio, Solon, OH; GAPDH (FL-335), iPLA2β (sc-14,463), tubulin (sc-166,729), and 2 ◦ (sc-2004, sc-2418, sc-2302) antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA; dispase (D4693), protease inhibitor cocktail (P8340), common reagents from Sigma Chemical Co., St. Louis, MO; and alpha-MEM media (12,561,049), collagenase 2 (17101− 015), enhanced chemiluminescence reagent (34,095), fetal bovine serum premium (FBS, S11150), and common cell culture reagents from ThermoFisher, Waltham, MA; The EP4 receptor agonist ONO-AE1− 329 and EP4 receptor antagonist ONO-AE3− 208 were kind gifts from ONO Pharmaceuticals, Osaka, Japan. .. (A) Bright field microscopy was performed with an Olympus IX81 microscope; (B) von Kossa staining was performed on fixed cells with a 5% silver nitrate solution [37], rinsed well, and exposed to UV light; and (C) alizarin red staining was performed on fixed cells with a 2% alizarin red solution [38], rinsed well, and scanned for imaging, and quantified using Image J software (National Institutes of Health).
Cell Culture:Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t
Article Snippet: .. Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards, and Triton X-100 (161-0324, 161-0407; BioRad, Hercules, CA); S -BEL (iPLA 2 β inhibitor), PGE 2 (10006801, 14010; Cayman Chemicals, Ann Arbor, MI); acetyl histone H4 (AcH4antibody, normal mouse IgG, protein A beads (06-911, 12-371, 16-125; EMD Millipore, Billerica, MA); Collagenase 2, fetal bovine serum (FBS) and common cell culture reagents (17101-015; Gibco, Carlsbad, CA); Immobilin-P PVDF membrane (IPVH00010; Millipore Corp., Bedford, MA; protease inhibitor cocktail (D4693), alpha-MEM media (P8340), and common reagents (Sigma Chemical Co., St. Louis, MO); and enhanced chemiluminescence reagent, (34095; Thermo-Fisher, Waltham, MA). .. Rat-derived INS-1 β-cells were cultured in Roswell Park Memorial Institute (RPMI, Cat. 11875093, Thermo Fisher Scientific Inc. Waltham, MA ) 1640 medium, containing 11 mM glucose, 10% fetal calf serum, 10 mM HEPES buffer, 2 mM glutamine, 1 mM sodium pyruvate, 50 mM β-mercaptoethanol (BME), and 0.1% (w/v) each of penicillin and streptomycin in cell culture conditions (37°C, 5%CO 2 /95% air).
Article Title: Ca 2+ -independent phospholipase A 2 β-derived PGE 2 contributes to osteogenesis.
Article Snippet: .. Reagents used in our studies were obtained from the following sources (catalog numbers): Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards (161− 0324), and Triton X-100 (161− 0407) from BioRad, Hercules, CA; S-BEL (iPLA2β inhibitor, 10,006,801), R-BEL (iPLA2γ inhibitor, 10,006,800), CAY10502 (cPLA2α inhibitor, 10,008,657), PGE2 (14,010), and PGE2 EIA Kit (514,010) from Cayman Chemicals, Ann Arbor, MI); paraformaldehyde from Electron Microscopy Sciences, Ft. Washington, PA; acetyl histone H3 antibody (06–911), normal mouse IgG (12–371), protein A beads (16–125) from EMD Millipore, Billerica, MA; Immobilon-P PVDF membrane (IPVH00010) from Millipore Corp., Bedford, MA; Runx2 antibody (D130− 3) from MBL International, Wobrurn, MA); arachidonic acid (ICN 19,462,510) from MP Bio, Solon, OH; GAPDH (FL-335), iPLA2β (sc-14,463), tubulin (sc-166,729), and 2 ◦ (sc-2004, sc-2418, sc-2302) antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA; dispase (D4693), protease inhibitor cocktail (P8340), common reagents from Sigma Chemical Co., St. Louis, MO; and alpha-MEM media (12,561,049), collagenase 2 (17101− 015), enhanced chemiluminescence reagent (34,095), fetal bovine serum premium (FBS, S11150), and common cell culture reagents from ThermoFisher, Waltham, MA; The EP4 receptor agonist ONO-AE1− 329 and EP4 receptor antagonist ONO-AE3− 208 were kind gifts from ONO Pharmaceuticals, Osaka, Japan. .. (A) Bright field microscopy was performed with an Olympus IX81 microscope; (B) von Kossa staining was performed on fixed cells with a 5% silver nitrate solution [37], rinsed well, and exposed to UV light; and (C) alizarin red staining was performed on fixed cells with a 2% alizarin red solution [38], rinsed well, and scanned for imaging, and quantified using Image J software (National Institutes of Health).
Membrane:Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t
Article Snippet: .. Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards, and Triton X-100 (161-0324, 161-0407; BioRad, Hercules, CA); S -BEL (iPLA 2 β inhibitor), PGE 2 (10006801, 14010; Cayman Chemicals, Ann Arbor, MI); acetyl histone H4 (AcH4antibody, normal mouse IgG, protein A beads (06-911, 12-371, 16-125; EMD Millipore, Billerica, MA); Collagenase 2, fetal bovine serum (FBS) and common cell culture reagents (17101-015; Gibco, Carlsbad, CA); Immobilin-P PVDF membrane (IPVH00010; Millipore Corp., Bedford, MA; protease inhibitor cocktail (D4693), alpha-MEM media (P8340), and common reagents (Sigma Chemical Co., St. Louis, MO); and enhanced chemiluminescence reagent, (34095; Thermo-Fisher, Waltham, MA). .. Rat-derived INS-1 β-cells were cultured in Roswell Park Memorial Institute (RPMI, Cat. 11875093, Thermo Fisher Scientific Inc. Waltham, MA ) 1640 medium, containing 11 mM glucose, 10% fetal calf serum, 10 mM HEPES buffer, 2 mM glutamine, 1 mM sodium pyruvate, 50 mM β-mercaptoethanol (BME), and 0.1% (w/v) each of penicillin and streptomycin in cell culture conditions (37°C, 5%CO 2 /95% air).
Article Title: Ca 2+ -independent phospholipase A 2 β-derived PGE 2 contributes to osteogenesis.
Article Snippet: .. Reagents used in our studies were obtained from the following sources (catalog numbers): Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards (161− 0324), and Triton X-100 (161− 0407) from BioRad, Hercules, CA; S-BEL (iPLA2β inhibitor, 10,006,801), R-BEL (iPLA2γ inhibitor, 10,006,800), CAY10502 (cPLA2α inhibitor, 10,008,657), PGE2 (14,010), and PGE2 EIA Kit (514,010) from Cayman Chemicals, Ann Arbor, MI); paraformaldehyde from Electron Microscopy Sciences, Ft. Washington, PA; acetyl histone H3 antibody (06–911), normal mouse IgG (12–371), protein A beads (16–125) from EMD Millipore, Billerica, MA; Immobilon-P PVDF membrane (IPVH00010) from Millipore Corp., Bedford, MA; Runx2 antibody (D130− 3) from MBL International, Wobrurn, MA); arachidonic acid (ICN 19,462,510) from MP Bio, Solon, OH; GAPDH (FL-335), iPLA2β (sc-14,463), tubulin (sc-166,729), and 2 ◦ (sc-2004, sc-2418, sc-2302) antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA; dispase (D4693), protease inhibitor cocktail (P8340), common reagents from Sigma Chemical Co., St. Louis, MO; and alpha-MEM media (12,561,049), collagenase 2 (17101− 015), enhanced chemiluminescence reagent (34,095), fetal bovine serum premium (FBS, S11150), and common cell culture reagents from ThermoFisher, Waltham, MA; The EP4 receptor agonist ONO-AE1− 329 and EP4 receptor antagonist ONO-AE3− 208 were kind gifts from ONO Pharmaceuticals, Osaka, Japan. .. (A) Bright field microscopy was performed with an Olympus IX81 microscope; (B) von Kossa staining was performed on fixed cells with a 5% silver nitrate solution [37], rinsed well, and exposed to UV light; and (C) alizarin red staining was performed on fixed cells with a 2% alizarin red solution [38], rinsed well, and scanned for imaging, and quantified using Image J software (National Institutes of Health).
Protease Inhibitor:Article Title: ER Stress-Induced β-Cell Apoptosis is Linked to Novel Select Lipid Signaling at the Transcriptional Level: Implications in T1D Developmen t
Article Snippet: .. Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards, and Triton X-100 (161-0324, 161-0407; BioRad, Hercules, CA); S -BEL (iPLA 2 β inhibitor), PGE 2 (10006801, 14010; Cayman Chemicals, Ann Arbor, MI); acetyl histone H4 (AcH4antibody, normal mouse IgG, protein A beads (06-911, 12-371, 16-125; EMD Millipore, Billerica, MA); Collagenase 2, fetal bovine serum (FBS) and common cell culture reagents (17101-015; Gibco, Carlsbad, CA); Immobilin-P PVDF membrane (IPVH00010; Millipore Corp., Bedford, MA; protease inhibitor cocktail (D4693), alpha-MEM media (P8340), and common reagents (Sigma Chemical Co., St. Louis, MO); and enhanced chemiluminescence reagent, (34095; Thermo-Fisher, Waltham, MA). .. Rat-derived INS-1 β-cells were cultured in Roswell Park Memorial Institute (RPMI, Cat. 11875093, Thermo Fisher Scientific Inc. Waltham, MA ) 1640 medium, containing 11 mM glucose, 10% fetal calf serum, 10 mM HEPES buffer, 2 mM glutamine, 1 mM sodium pyruvate, 50 mM β-mercaptoethanol (BME), and 0.1% (w/v) each of penicillin and streptomycin in cell culture conditions (37°C, 5%CO 2 /95% air).
Article Title: Ca 2+ -independent phospholipase A 2 β-derived PGE 2 contributes to osteogenesis.
Article Snippet: .. Reagents used in our studies were obtained from the following sources (catalog numbers): Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards (161− 0324), and Triton X-100 (161− 0407) from BioRad, Hercules, CA; S-BEL (iPLA2β inhibitor, 10,006,801), R-BEL (iPLA2γ inhibitor, 10,006,800), CAY10502 (cPLA2α inhibitor, 10,008,657), PGE2 (14,010), and PGE2 EIA Kit (514,010) from Cayman Chemicals, Ann Arbor, MI); paraformaldehyde from Electron Microscopy Sciences, Ft. Washington, PA; acetyl histone H3 antibody (06–911), normal mouse IgG (12–371), protein A beads (16–125) from EMD Millipore, Billerica, MA; Immobilon-P PVDF membrane (IPVH00010) from Millipore Corp., Bedford, MA; Runx2 antibody (D130− 3) from MBL International, Wobrurn, MA); arachidonic acid (ICN 19,462,510) from MP Bio, Solon, OH; GAPDH (FL-335), iPLA2β (sc-14,463), tubulin (sc-166,729), and 2 ◦ (sc-2004, sc-2418, sc-2302) antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA; dispase (D4693), protease inhibitor cocktail (P8340), common reagents from Sigma Chemical Co., St. Louis, MO; and alpha-MEM media (12,561,049), collagenase 2 (17101− 015), enhanced chemiluminescence reagent (34,095), fetal bovine serum premium (FBS, S11150), and common cell culture reagents from ThermoFisher, Waltham, MA; The EP4 receptor agonist ONO-AE1− 329 and EP4 receptor antagonist ONO-AE3− 208 were kind gifts from ONO Pharmaceuticals, Osaka, Japan. .. (A) Bright field microscopy was performed with an Olympus IX81 microscope; (B) von Kossa staining was performed on fixed cells with a 5% silver nitrate solution [37], rinsed well, and exposed to UV light; and (C) alizarin red staining was performed on fixed cells with a 2% alizarin red solution [38], rinsed well, and scanned for imaging, and quantified using Image J software (National Institutes of Health).
Enzyme-linked Immunosorbent Assay:Article Title: Ca 2+ -independent phospholipase A 2 β-derived PGE 2 contributes to osteogenesis.
Article Snippet: .. Reagents used in our studies were obtained from the following sources (catalog numbers): Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards (161− 0324), and Triton X-100 (161− 0407) from BioRad, Hercules, CA; S-BEL (iPLA2β inhibitor, 10,006,801), R-BEL (iPLA2γ inhibitor, 10,006,800), CAY10502 (cPLA2α inhibitor, 10,008,657), PGE2 (14,010), and PGE2 EIA Kit (514,010) from Cayman Chemicals, Ann Arbor, MI); paraformaldehyde from Electron Microscopy Sciences, Ft. Washington, PA; acetyl histone H3 antibody (06–911), normal mouse IgG (12–371), protein A beads (16–125) from EMD Millipore, Billerica, MA; Immobilon-P PVDF membrane (IPVH00010) from Millipore Corp., Bedford, MA; Runx2 antibody (D130− 3) from MBL International, Wobrurn, MA); arachidonic acid (ICN 19,462,510) from MP Bio, Solon, OH; GAPDH (FL-335), iPLA2β (sc-14,463), tubulin (sc-166,729), and 2 ◦ (sc-2004, sc-2418, sc-2302) antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA; dispase (D4693), protease inhibitor cocktail (P8340), common reagents from Sigma Chemical Co., St. Louis, MO; and alpha-MEM media (12,561,049), collagenase 2 (17101− 015), enhanced chemiluminescence reagent (34,095), fetal bovine serum premium (FBS, S11150), and common cell culture reagents from ThermoFisher, Waltham, MA; The EP4 receptor agonist ONO-AE1− 329 and EP4 receptor antagonist ONO-AE3− 208 were kind gifts from ONO Pharmaceuticals, Osaka, Japan. .. (A) Bright field microscopy was performed with an Olympus IX81 microscope; (B) von Kossa staining was performed on fixed cells with a 5% silver nitrate solution [37], rinsed well, and exposed to UV light; and (C) alizarin red staining was performed on fixed cells with a 2% alizarin red solution [38], rinsed well, and scanned for imaging, and quantified using Image J software (National Institutes of Health).
Electron Microscopy:Article Title: Ca 2+ -independent phospholipase A 2 β-derived PGE 2 contributes to osteogenesis.
Article Snippet: .. Reagents used in our studies were obtained from the following sources (catalog numbers): Coomassie reagents, SDS-PAGE reagents, kaleidoscope pre-stained molecular mass standards (161− 0324), and Triton X-100 (161− 0407) from BioRad, Hercules, CA; S-BEL (iPLA2β inhibitor, 10,006,801), R-BEL (iPLA2γ inhibitor, 10,006,800), CAY10502 (cPLA2α inhibitor, 10,008,657), PGE2 (14,010), and PGE2 EIA Kit (514,010) from Cayman Chemicals, Ann Arbor, MI); paraformaldehyde from Electron Microscopy Sciences, Ft. Washington, PA; acetyl histone H3 antibody (06–911), normal mouse IgG (12–371), protein A beads (16–125) from EMD Millipore, Billerica, MA; Immobilon-P PVDF membrane (IPVH00010) from Millipore Corp., Bedford, MA; Runx2 antibody (D130− 3) from MBL International, Wobrurn, MA); arachidonic acid (ICN 19,462,510) from MP Bio, Solon, OH; GAPDH (FL-335), iPLA2β (sc-14,463), tubulin (sc-166,729), and 2 ◦ (sc-2004, sc-2418, sc-2302) antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA; dispase (D4693), protease inhibitor cocktail (P8340), common reagents from Sigma Chemical Co., St. Louis, MO; and alpha-MEM media (12,561,049), collagenase 2 (17101− 015), enhanced chemiluminescence reagent (34,095), fetal bovine serum premium (FBS, S11150), and common cell culture reagents from ThermoFisher, Waltham, MA; The EP4 receptor agonist ONO-AE1− 329 and EP4 receptor antagonist ONO-AE3− 208 were kind gifts from ONO Pharmaceuticals, Osaka, Japan. .. (A) Bright field microscopy was performed with an Olympus IX81 microscope; (B) von Kossa staining was performed on fixed cells with a 5% silver nitrate solution [37], rinsed well, and exposed to UV light; and (C) alizarin red staining was performed on fixed cells with a 2% alizarin red solution [38], rinsed well, and scanned for imaging, and quantified using Image J software (National Institutes of Health).
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