Azithromycin (CP 62993) is a macrolide antibiotic that primarily targets the 50S subunit of bacterial ribosomes. Azithromycin is commonly used in studies of bacterial infections.

PBMCs proliferation:5 µg/mL, IL-1β:17.4 μM, TNF-α:10.66 μM, Parasite growth:8.4 ± 1.2 μM, NF-κB:56 μM (A549 cells)

THP-1 cells (106 cells in 1 mL RPMI medium, without antibiotics, growth factors or serum) are seeded in each well of 24-well plates and allowed to settle for 1 hour. Next, 50 μL of the test compound is added followed by 50 μL of LPS (final concentration of 10 μg/mL). After 24h (37°C and 5% CO2) the supernatants and cell pellets are collected (1200 rpm, 5 min). THP-1 cell viability is tested using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). MTT is dissolved at 2 mg/mL in PBS and aliquots are stored at -20°C. The MTT assay is performed according to the suppliers instructions. Absorbance of MTT converted into formazan is measured at a wavelength of 570 nm with background subtraction at 630 nm.