Biomarker Discovery:Article Title: Integrative multi-omics landscape of fluoxetine action across 27 brain regions reveals global increase in energy metabolism and region-specific chromatin remodelling
Article Snippet: .. Validation of Mitochondrial and Oxphos trends: a) Cell culture C6 rat glioma cell line (ATCC CRL-2199, RRID:CVCL_5657) was cultured in DMEM (4.5 g/L glucose) (Gibco, Cat# 11995065) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, Cat# 10082147) and incubated at 37°C with 5% CO2 in a humidified atmosphere. b) Drug treatment C6 rat glioma cells were seeded at a density of 50,000 cells into T25 flasks and treated with fluoxetine (Sigma, Cat# F132) after overnight incubation at 37°C with 5% CO2 in a humidified atmosphere. .. 6 h after addition of fluoxetine, either corticosterone (Sigma, Cat# 27840) or ethanol was added for the respective treatment groups and incubated for another 66 h (total 72 h). c) Mito stress assay After 72 h drug treatment, cells in all treatment groups were harvested and seeded at a density of 30,000 cells (in respective treatment media) into each well of a Seahorse XF96 V3 PS microplate (Agilent Technologies, Cat# 101085-004) for the mito stress assay and a 96-well black, clear bottom plate (Corning, Cat# 3904) for cell number normalisation.
Cell Culture:Article Title: Integrative multi-omics landscape of fluoxetine action across 27 brain regions reveals global increase in energy metabolism and region-specific chromatin remodelling
Article Snippet: .. Validation of Mitochondrial and Oxphos trends: a) Cell culture C6 rat glioma cell line (ATCC CRL-2199, RRID:CVCL_5657) was cultured in DMEM (4.5 g/L glucose) (Gibco, Cat# 11995065) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, Cat# 10082147) and incubated at 37°C with 5% CO2 in a humidified atmosphere. b) Drug treatment C6 rat glioma cells were seeded at a density of 50,000 cells into T25 flasks and treated with fluoxetine (Sigma, Cat# F132) after overnight incubation at 37°C with 5% CO2 in a humidified atmosphere. .. 6 h after addition of fluoxetine, either corticosterone (Sigma, Cat# 27840) or ethanol was added for the respective treatment groups and incubated for another 66 h (total 72 h). c) Mito stress assay After 72 h drug treatment, cells in all treatment groups were harvested and seeded at a density of 30,000 cells (in respective treatment media) into each well of a Seahorse XF96 V3 PS microplate (Agilent Technologies, Cat# 101085-004) for the mito stress assay and a 96-well black, clear bottom plate (Corning, Cat# 3904) for cell number normalisation.
Incubation:Article Title: Integrative multi-omics landscape of fluoxetine action across 27 brain regions reveals global increase in energy metabolism and region-specific chromatin remodelling
Article Snippet: .. Validation of Mitochondrial and Oxphos trends: a) Cell culture C6 rat glioma cell line (ATCC CRL-2199, RRID:CVCL_5657) was cultured in DMEM (4.5 g/L glucose) (Gibco, Cat# 11995065) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, Cat# 10082147) and incubated at 37°C with 5% CO2 in a humidified atmosphere. b) Drug treatment C6 rat glioma cells were seeded at a density of 50,000 cells into T25 flasks and treated with fluoxetine (Sigma, Cat# F132) after overnight incubation at 37°C with 5% CO2 in a humidified atmosphere. .. 6 h after addition of fluoxetine, either corticosterone (Sigma, Cat# 27840) or ethanol was added for the respective treatment groups and incubated for another 66 h (total 72 h). c) Mito stress assay After 72 h drug treatment, cells in all treatment groups were harvested and seeded at a density of 30,000 cells (in respective treatment media) into each well of a Seahorse XF96 V3 PS microplate (Agilent Technologies, Cat# 101085-004) for the mito stress assay and a 96-well black, clear bottom plate (Corning, Cat# 3904) for cell number normalisation.
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