Luciferin wehd substrate

For caspase-1 and LDH determination, 2 × 10⁵ human CD14⁺ monocytes were plated on 48-well plates in RPMI 10% FBS and incubated overnight. The following day, cells were infected with SARS-CoV-2 using MOI 5 in RPMI without Phenol Red (3.5 g/liter Hepes, 2 g/liter NaHCO₃, 10.4 g/liter RPMI without Phenol Red, and 1% glutamine, pH 7.2). After 1 h of virus adsorption, fresh media (RPMI 2% FBS without Phenol Red) with or without MCC950 at 10 µM were added, and cells were incubated for 24 h. To measure caspase-1 activity, the supernatants were collected and incubated with the Luciferin WEHD-substrate provided by the Caspase-Glo 1 Assay (Promega). After 1 h of incubation at room temperature, luminescence was measured using the SpectraMax i3 system (Molecular Devices). LDH release was measured in the supernatants using the CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega) following the manufacturer’s instructions.

To measure caspase-1 activity, the supernatants were collected and incubated with the Luciferin WEHD substrate provided by the Caspase-Glo 1 assay (Promega, WI, USA). After 1 hour of incubation at room temperature, luminescence was measured using SpectraMax i3x Multi-Mode Microplate (Molecular Devices).