2.3. Luciferase Reporter Assay

Reporter cells (RAW-NF-κB-Luc, RAW-NFAT-Luc, and RAW-AP-1-Luc) were cultured in DMEM medium (GE Healthcare, USA) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, USA), 50 U/mL penicillin, 50 μg/mL streptomycin, 2 mM glutamine, and 0.1 M NaHCO₃ (all PanEco, Russia) at 37°C with 5% CO₂. We used the protocol described in our previous work [16]; namely, experiment cells were seeded at 1 × 10⁵ cells/well in 96-well plates (100 μL/well). The next day, C-type lectin receptor agonists were added to the wells to a final concentration of 20 μg/mL, 4 μg/mL, and 1 μg/mL. Eight hours later, to detect luciferase activity, 100 μL of Bright-Glo Luciferase Assay Buffer containing luciferin substrate (Promega, USA) was added to each well. Luminescence was measured in relative units using a Synergy H4 Hybrid Reader (BioTek, Germany).