Reverse Transcription:Article Title: Ventromedial hypothalamic nucleus neuronal nitric oxide knockdown effects on GABAergic neuron metabolic sensor and transmitter marker gene expression in the male rat
Article Snippet: Lysate RNA quantity and purity were verified by NanoDrop spectrophotometry (prod. no. ND-ONE-W, ThermoFisherScientific, Waltham, MA). .. RNA samples were reverse-transcribed to cDNA by addition of iScriptTM Advanced cDNA Synthesis kit reagents (prod. no. 1725038; Bio-Rad) and centrifugation at 46 °C (20 min) and 95 °C (1 min), as reported [ – ]. .. A preamplification master mix was prepared by combining a pool of 15 PrimePCRTM PreAmps for SYBR ® Green Assays [GAD1/GAD67 (prod. no. qRnoCID0004554), GAD2/GAD65 (prod. no. qRnoCID0003485), nNOS/NOS1 (prod. no. qRnoCED0009301), Ghrh (prod. no. qRnoCID0007723), SF-1/Nr5a1 (prod. no. qRnoCID0001458), GPR81/HCAR1 (prod. no. qRnoCED0001040), GCK (prod. no. qRnoCID0009394), AMPKα1/PRKAA1 (prod. no. qRnoCID0001262), AMPKα1/PRKAA2 (prod. no. qRnoCID0006799), excitatory amino acid transporter-2 (EAAT-2/Slc1A2) (prod. no. qRnoCED0005967), GLUT2/Slc2A2 (prod. no. qRnoCID0009479), GLUT3/Slc2A3 (prod. no. qRnoCID0004167), housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (prod. no. qRnoCID0057018)] with SsoAdvancedTM PreAmp Supermix (prod. no. 1725160; Bio-Rad).
Article Title: In Vivo Glucose Transporter-2 Regulation of Dorsomedial Versus Ventrolateral VMN Astrocyte Metabolic Sensor and Glycogen Metabolic Enzyme Gene Expression in Female Rat.
Article Snippet: Astrocyte glycogenolysis shapes ventromedial hypothalamic nucleus (VMN) regulation of glucostasis in vivo.. Glucose transporter-2 (GLUT2), a plasma membrane glucose sensor, controls hypothalamic primary astrocyte culture glycogen metabolism in vitro.. In vivo gene silencing tools and single-cell laser-catapult-microdissection/multiplex qPCR techniques were used here to examine whether GLUT2 governs dorsomedial (VMNdm) and/or ventrolateral (VMNvl) VMN astrocyte metabolic sensor and glycogen metabolic enzyme gene profiles.
Article Title: Glucose transporter-2 regulation of VMN GABA neuron metabolic sensor and transmitter gene expression
Article Snippet: Complementary DNA (cDNA) Synthesis and Amplification : Individual neurons collected into lysis buffer were sequentially incubated at 25 °C (10 min), then 75 °C (5 min) in a Bio-Rad iCyclerQ RT-PCR Detection System. .. RNA samples were reverse-transcribed to cDNA using iScriptTM Advanced cDNA Synthesis kit reagents (prod. no. 1725038; Bio-Rad), using described methods , , . .. Preamplification master mix was prepared by combining Bio-Rad PrimePCRTM PreAmps for SYBR® Green Assays for GAD1/GAD67 (prod. no. qRnoCID0004554), GAD2/GAD65 (prod. no. qRnoCID0003485), NOS1/nNOS (prod. no. qRnoCED0009301), Ghrh (prod. no. qRnoCID0007723), Slc1A2/GLT-1 (prod. no. qRnoCED0005967), PRKAA1 (prod. no. qRnoCID0001262), PRKAA2 (prod. no. qRnoCID0006799), Nr5a1/SF-1 (prod. no. qRnoCID0001458), Slc2A2 (prod. no. qRnoCID0009479), and GAPDH (prod. no. qRnoCID0057018) with SsoAdvancedTM PreAmp Supermix (prod. no. 1725160 Bio-Rad ).
Article Title: Ventromedial hypothalamic nucleus neuronal nitric oxide knockdown effects on GABAergic neuron metabolic sensor and transmitter marker gene expression in the male rat.
Article Snippet: Lysate RNA quantity and purity were verified by NanoDrop spectrophotometry (prod. no. ND-ONE-W, ThermoFisherScientific, Waltham, MA). .. RNA samples were reverse-transcribed to cDNA by addition of iScriptTM Advanced cDNA Synthesis kit reagents (prod. no. 1725038; Bio-Rad) and centrifugation at 46 °C (20 min) and 95 °C (1 min), as reported [29–31]. .. A preamplification master mix was prepared by combining a pool of 15 PrimePCRTM PreAmps for SYBR® Green Assays [GAD1/GAD67 (prod. no. qRnoCID0004554), GAD2/ GAD65 (prod. no. qRnoCID0003485), nNOS/NOS1 (prod. no. qRnoCED0009301), Ghrh (prod. no. qRnoCID0007723), SF-1/Nr5a1 (prod. no. qRnoCID0001458), GPR81/HCAR1 (prod. no. qRnoCED0001040), GCK (prod. no. qRnoCID0009394), AMPKα1/PRKAA1 (prod. no. qRnoCID0001262), AMPKα1/PRKAA2 (prod. no. qRnoCID0006799), excitatory amino acid transporter-2 (EAAT-2/Slc1A2) (prod. no. qRnoCED0005967), GLUT2/Slc2A2 (prod. no. qRnoCID0009479), GLUT3/ Slc2A3 (prod. no. qRnoCID0004167), housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (prod. no. qRnoCID0057018)] with SsoAdvancedTM PreAmp Supermix (prod. no. 1725160; Bio-Rad).
cDNA Synthesis:Article Title: Ventromedial hypothalamic nucleus neuronal nitric oxide knockdown effects on GABAergic neuron metabolic sensor and transmitter marker gene expression in the male rat
Article Snippet: Lysate RNA quantity and purity were verified by NanoDrop spectrophotometry (prod. no. ND-ONE-W, ThermoFisherScientific, Waltham, MA). .. RNA samples were reverse-transcribed to cDNA by addition of iScriptTM Advanced cDNA Synthesis kit reagents (prod. no. 1725038; Bio-Rad) and centrifugation at 46 °C (20 min) and 95 °C (1 min), as reported [ – ]. .. A preamplification master mix was prepared by combining a pool of 15 PrimePCRTM PreAmps for SYBR ® Green Assays [GAD1/GAD67 (prod. no. qRnoCID0004554), GAD2/GAD65 (prod. no. qRnoCID0003485), nNOS/NOS1 (prod. no. qRnoCED0009301), Ghrh (prod. no. qRnoCID0007723), SF-1/Nr5a1 (prod. no. qRnoCID0001458), GPR81/HCAR1 (prod. no. qRnoCED0001040), GCK (prod. no. qRnoCID0009394), AMPKα1/PRKAA1 (prod. no. qRnoCID0001262), AMPKα1/PRKAA2 (prod. no. qRnoCID0006799), excitatory amino acid transporter-2 (EAAT-2/Slc1A2) (prod. no. qRnoCED0005967), GLUT2/Slc2A2 (prod. no. qRnoCID0009479), GLUT3/Slc2A3 (prod. no. qRnoCID0004167), housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (prod. no. qRnoCID0057018)] with SsoAdvancedTM PreAmp Supermix (prod. no. 1725160; Bio-Rad).
Article Title: In Vivo Glucose Transporter-2 Regulation of Dorsomedial Versus Ventrolateral VMN Astrocyte Metabolic Sensor and Glycogen Metabolic Enzyme Gene Expression in Female Rat.
Article Snippet: Astrocyte glycogenolysis shapes ventromedial hypothalamic nucleus (VMN) regulation of glucostasis in vivo.. Glucose transporter-2 (GLUT2), a plasma membrane glucose sensor, controls hypothalamic primary astrocyte culture glycogen metabolism in vitro.. In vivo gene silencing tools and single-cell laser-catapult-microdissection/multiplex qPCR techniques were used here to examine whether GLUT2 governs dorsomedial (VMNdm) and/or ventrolateral (VMNvl) VMN astrocyte metabolic sensor and glycogen metabolic enzyme gene profiles.
Article Title: Glucose transporter-2 regulation of VMN GABA neuron metabolic sensor and transmitter gene expression
Article Snippet: Complementary DNA (cDNA) Synthesis and Amplification : Individual neurons collected into lysis buffer were sequentially incubated at 25 °C (10 min), then 75 °C (5 min) in a Bio-Rad iCyclerQ RT-PCR Detection System. .. RNA samples were reverse-transcribed to cDNA using iScriptTM Advanced cDNA Synthesis kit reagents (prod. no. 1725038; Bio-Rad), using described methods , , . .. Preamplification master mix was prepared by combining Bio-Rad PrimePCRTM PreAmps for SYBR® Green Assays for GAD1/GAD67 (prod. no. qRnoCID0004554), GAD2/GAD65 (prod. no. qRnoCID0003485), NOS1/nNOS (prod. no. qRnoCED0009301), Ghrh (prod. no. qRnoCID0007723), Slc1A2/GLT-1 (prod. no. qRnoCED0005967), PRKAA1 (prod. no. qRnoCID0001262), PRKAA2 (prod. no. qRnoCID0006799), Nr5a1/SF-1 (prod. no. qRnoCID0001458), Slc2A2 (prod. no. qRnoCID0009479), and GAPDH (prod. no. qRnoCID0057018) with SsoAdvancedTM PreAmp Supermix (prod. no. 1725160 Bio-Rad ).
Article Title: Ventromedial hypothalamic nucleus neuronal nitric oxide knockdown effects on GABAergic neuron metabolic sensor and transmitter marker gene expression in the male rat.
Article Snippet: Lysate RNA quantity and purity were verified by NanoDrop spectrophotometry (prod. no. ND-ONE-W, ThermoFisherScientific, Waltham, MA). .. RNA samples were reverse-transcribed to cDNA by addition of iScriptTM Advanced cDNA Synthesis kit reagents (prod. no. 1725038; Bio-Rad) and centrifugation at 46 °C (20 min) and 95 °C (1 min), as reported [29–31]. .. A preamplification master mix was prepared by combining a pool of 15 PrimePCRTM PreAmps for SYBR® Green Assays [GAD1/GAD67 (prod. no. qRnoCID0004554), GAD2/ GAD65 (prod. no. qRnoCID0003485), nNOS/NOS1 (prod. no. qRnoCED0009301), Ghrh (prod. no. qRnoCID0007723), SF-1/Nr5a1 (prod. no. qRnoCID0001458), GPR81/HCAR1 (prod. no. qRnoCED0001040), GCK (prod. no. qRnoCID0009394), AMPKα1/PRKAA1 (prod. no. qRnoCID0001262), AMPKα1/PRKAA2 (prod. no. qRnoCID0006799), excitatory amino acid transporter-2 (EAAT-2/Slc1A2) (prod. no. qRnoCED0005967), GLUT2/Slc2A2 (prod. no. qRnoCID0009479), GLUT3/ Slc2A3 (prod. no. qRnoCID0004167), housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (prod. no. qRnoCID0057018)] with SsoAdvancedTM PreAmp Supermix (prod. no. 1725160; Bio-Rad).
Centrifugation:Article Title: Ventromedial hypothalamic nucleus neuronal nitric oxide knockdown effects on GABAergic neuron metabolic sensor and transmitter marker gene expression in the male rat
Article Snippet: Lysate RNA quantity and purity were verified by NanoDrop spectrophotometry (prod. no. ND-ONE-W, ThermoFisherScientific, Waltham, MA). .. RNA samples were reverse-transcribed to cDNA by addition of iScriptTM Advanced cDNA Synthesis kit reagents (prod. no. 1725038; Bio-Rad) and centrifugation at 46 °C (20 min) and 95 °C (1 min), as reported [ – ]. .. A preamplification master mix was prepared by combining a pool of 15 PrimePCRTM PreAmps for SYBR ® Green Assays [GAD1/GAD67 (prod. no. qRnoCID0004554), GAD2/GAD65 (prod. no. qRnoCID0003485), nNOS/NOS1 (prod. no. qRnoCED0009301), Ghrh (prod. no. qRnoCID0007723), SF-1/Nr5a1 (prod. no. qRnoCID0001458), GPR81/HCAR1 (prod. no. qRnoCED0001040), GCK (prod. no. qRnoCID0009394), AMPKα1/PRKAA1 (prod. no. qRnoCID0001262), AMPKα1/PRKAA2 (prod. no. qRnoCID0006799), excitatory amino acid transporter-2 (EAAT-2/Slc1A2) (prod. no. qRnoCED0005967), GLUT2/Slc2A2 (prod. no. qRnoCID0009479), GLUT3/Slc2A3 (prod. no. qRnoCID0004167), housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (prod. no. qRnoCID0057018)] with SsoAdvancedTM PreAmp Supermix (prod. no. 1725160; Bio-Rad).
Article Title: Ventromedial hypothalamic nucleus neuronal nitric oxide knockdown effects on GABAergic neuron metabolic sensor and transmitter marker gene expression in the male rat.
Article Snippet: Lysate RNA quantity and purity were verified by NanoDrop spectrophotometry (prod. no. ND-ONE-W, ThermoFisherScientific, Waltham, MA). .. RNA samples were reverse-transcribed to cDNA by addition of iScriptTM Advanced cDNA Synthesis kit reagents (prod. no. 1725038; Bio-Rad) and centrifugation at 46 °C (20 min) and 95 °C (1 min), as reported [29–31]. .. A preamplification master mix was prepared by combining a pool of 15 PrimePCRTM PreAmps for SYBR® Green Assays [GAD1/GAD67 (prod. no. qRnoCID0004554), GAD2/ GAD65 (prod. no. qRnoCID0003485), nNOS/NOS1 (prod. no. qRnoCED0009301), Ghrh (prod. no. qRnoCID0007723), SF-1/Nr5a1 (prod. no. qRnoCID0001458), GPR81/HCAR1 (prod. no. qRnoCED0001040), GCK (prod. no. qRnoCID0009394), AMPKα1/PRKAA1 (prod. no. qRnoCID0001262), AMPKα1/PRKAA2 (prod. no. qRnoCID0006799), excitatory amino acid transporter-2 (EAAT-2/Slc1A2) (prod. no. qRnoCED0005967), GLUT2/Slc2A2 (prod. no. qRnoCID0009479), GLUT3/ Slc2A3 (prod. no. qRnoCID0004167), housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (prod. no. qRnoCID0057018)] with SsoAdvancedTM PreAmp Supermix (prod. no. 1725160; Bio-Rad).
|