Huh7 cells were plated in white walled 96-well plates (Corning, Lowell, MA) and incubated with each compound in 2-fold serial dilutions. After 1 h, VSVΔG-EBOV-GP or VSVΔG-VSV-G was added. The cells were challenged with virus in the presence of compounds at 37 °C for 16 h, and the medium was replaced with luciferase assay buffer (20 mM Tricine-HCl, pH 7.5, 8 mM MgSO4, 0.13 mM ethylenediaminetetraacetic acid (EDTA), 0.53 mM ATP, 33 mM dithiothreitol (DTT) 0.47 mM luciferin) containing 0.2% of Triton X-100 detergent. After the cells were incubated with the buffer for 10 min at room temperature, the luciferase activity was measured using a 96-well plate luminometer (Promega).
96 well plate luminometer
Manufactured by Promega
Sourced in United States
The 96-well plate luminometer is a laboratory instrument designed to measure luminescence from samples in a 96-well microplate format. It is used to quantify the level of light emitted from luciferase-based assays or other luminescent reactions within the wells of a microplate.
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6 protocols using 96 well plate luminometer
1
Virus Inhibition Assay in Huh7 Cells
2
Cell Viability Assay for Huh7 Cells
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Drug cytotoxicity was measured using CellTiter-Glo Luminescent Cell Viability Assay (Promega, Madison, WI) following the manufacturer's protocol. Huh7 cells were seeded in white walled 96 well plates (Corning, Lowell, MA) and incubated with each compound at 37 °C. After 24 h, the assay buffer was added to the culture plates and incubated for additional 10 min at room temperature. Luminescence was measured using a 96-well plate luminometer (Promega, Madison, WI).
3
Luciferase-based EBOV entry assay
4
CellTiter-Glo Cell Viability Assay
5
Innate Immune Signaling Pathway Activation
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HEK293 cells were co-transfected with the IFN-β promoter, ISRE promoter and NF-kB promoter luciferase reporter plasmid and a TK-Renilla luciferase reporter, together with vector alone or USP12, USP12C48A, IFI16, RIG-I, MAVS, IRF3-5D, IRF7, cGAS, STING, TBK1, TLR9, MyD88, and DDX41 constructs. Twenty-four hours later, cells were infected with HSV or VSV for 6 h. Luciferase reporter activities were measured in triplicate using the Dual-Luciferase reporter assay system (Promega, Madison, WI, USA), according to the manufacturer’s protocol, and quantified using the 96-well plate luminometer (Promega). The firefly luciferase to Renilla luciferase ratios were determined and were defined as the relative luciferase activity.
6
Endocytosis Measurement of Secreted Proteins
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Cells (90,000 per well) stably expressing LgBiT were plated in 24-well culture dishes. After 24 h, the medium was changed and cells were incubated for 30 min with the medium containing HiBiT fusions (taken from cells expressing Shh-HiBiT or secreted mCherry, SecCh-HiBiT for 48 h, and adjusted on protein Luciferase activity), before incubating the cells with trypsin and removing them. After centrifugation, cells were lysed and the luciferase activity of endocytosed protein was measured with a 96-well plate luminometer (Tristar, Berthold) with a HiBiT assay kit (Promega).
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