Cell viability was measured following manufacturer’s instructions of CellTiter-GloR Luminescent Cell Viability Assay (Promega). Caspase 3 activity was determined by measuring the cleavage of the fluorescent substrate, Acetyl-Asp-Glu-Val-Asp-7-amino-4-trifluoromrthylcoumarin (EMD Millipore). Cell lysates for the caspase 3 activities were prepared without protease inhibitors as previously described (Saikia et al., 2014 (link)). Proteasome activity was measured following manufacturer’s instructions of Proteasome-Glo™ Assay (Chymotrypsin-like) Systems (Promega).
Celltiter glor luminescent cell viability assay
Manufactured by Promega
Sourced in United States
The CellTiter-GloR Luminescent Cell Viability Assay is a quantitative, homogeneous method for determining the number of viable cells in a sample. The assay measures the amount of ATP present, which indicates the metabolically active cells. This is achieved through a luminescent reaction that produces light in the presence of ATP.
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5 protocols using celltiter glor luminescent cell viability assay
1
Cell Viability, Caspase 3, and Proteasome Assays
2
Cell Proliferation Assay Protocol
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For cell counting kit‐8 (CCK‐8), after 48 h of transfection, a total of approximately 2 × 103 cells/well was seeded in 96‐well plate. After culturing at indicated time (0, 1, 2, 3, and 4 day), the cellular proliferation was detected using CellTiter‐GloR Luminescent Cell Viability Assay (Promega, Madison, WI, USA) according to manufacturer's instructions.
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well, and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click‐iTR EdU Kit, and images were visualized using fluorescent microscope (Olympus).
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well, and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click‐iTR EdU Kit, and images were visualized using fluorescent microscope (Olympus).
3
Assessing GDF15-induced cell viability
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Cells were seeded in triplicate into a 96-well plate (5,000 cells/well) and incubated with EBM2 + 0.5% FBS for 7h. Then, cells were treated with 50 ng/mL of GDF15 overnight. Cell viability was assessed using the Cell titer gloR Luminescent cell viability assay (Promega) following the manufacturer’s instructions. Luminescence was measured using a Microplate Luminometer Centro LB 960 (Berthold Technologies).
4
Metabolic Profiling of Bone Marrow-Derived Macrophages
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Bone marrow-derived macrophages (BMDMs) were harvested and analyzed using the EnzyChrom NAD+/NADH Assay Kit (Bioassay Systems, Hayward, CA, USA), Elite NADPH Assay Kit (eEnzyme, Gaithersburg, MD, USA), and CellTiter-Glo(R) Luminescent Cell Viability assay (Promega, Madison, WI, USA) according to the manufacturers’ instructions.
5
Cell Proliferation Assays with CCK-8 and EdU
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For Cell Counting Kit-8 (CCK-8), after 48 h of transfection, a total of approximately 2 × 10 3 cells/well was seeded in 96-well plate. After culturing at indicated time (0, 6, 12, 24 and 48 days), the cellular proliferation was detected using CellTiter-GloR Luminescent Cell Viability Assay (Promega, Madison, WI) according to manufacturer's instructions.
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click-iTR EdU Kit and images were visualised using fluorescent microscope (Olympus).
For ethynyl deoxyuridine (EdU) incorporation assay, EdU (10 mM) was added to each well and cells were fixed with 4% formaldehyde for 30 min. After washing, EdU was detected with Click-iTR EdU Kit and images were visualised using fluorescent microscope (Olympus).
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