Cellometer vision cba analysis system

Mouse J774A.1 macrophages (ATCC, Rockville MD, USA) were maintained in DMEM supplemented with 10% FBS, 100 U/ml penicillin, and 100 µg/ml streptomycin at 37°C with 5% CO₂. Cells from passages six to nine were used in these studies. Human THP-1 monotypic cells (ATCC, Rockville MD, USA) were cultured in RPMI 1640 medium supplemented with 10% FBS, 100 U/ml penicillin, and 100 µg/ml streptomycin at 37°C with 5% CO₂. THP-1 monocytes were treated with PMA (100 ng/ml) for 5 days to facilitate differentiation into macrophages. HIV PIs (ritonavir and lopinavir) and Raltegravir were dissolved in dimethyl sulfoxide (DMSO) and added directly to culture medium (final concentrations 5 to 25 µM) and incubated for 0.5 to 24 h. For each result, a minimum of three independent experiments was performed. Cell viability was assessed with the Cellometer Vision CBA Analysis System (Nexcelom Bioscience, Lawrence, MA) and with the use of trypan blue.

SQ20B and FaDu cells were cultured in 6 well plates. After 24h treatment, cells were harvested and resuspended into a density of 10⁶cells/ml. BD Pharmingen FITC Annexin V Apoptosis Detection Kit I (BD Science, San Diego, CA) was used to perform the staining, according to the protocol recommended by the manufacturer as described previously [30 (link)]. The stained cells were then analyzed by Cellometer Vision CBA Analysis System (Nexcelom Bioscience, Lawrence, MA).