Murine macrophages (ATCC: TIB-67) and human lung epithelial cells (ATCC: HTB-177) were plated into 96 well plates and incubated overnight, then exposed to different concentrations of sodium molybdate (1, 3, 5 or 8 mM) or ce-MoS2 (up to 80 µg/mL) for 24, 48 or 72 hours. Cells were washed once, and then cell viability was assessed after 24, 48 and 72 hours using dehydrogenase activity assay Wst 8 (CCK-8; Dojindo; CK04-05). 10,000 lung epithelial cells or 25,000 macrophages were seeded and exposed, and the cells were washed once with cell culture medium and then exposed to 100 µl/well phenol red-free medium containing WST-8 substrate diluted to a concentration of 1:10 for one hour to lung epithelial cells or for 3 hours to murine macrophages. The absorbance was quantified at 450 nm using a Spectramax M2 (Molecular Devices).
Visualization of cell death in murine macrophages and human lung epithelial cells was conducted using ethidium-homodimer /Syto 10 stain after exposure to sodium molybdate (1, 3, 5 or 8 mM) or ce-MoS2 (up to 80 µg/mL) for 24 hours. Cells were washed once and incubated with 5 µM/ml Syto10/ and 1 µg/ml ethidium homodimer (Invitrogen) in phosphate buffered saline for 5 min (macrophages) or 15 min (lung epithelial cells). Images were visualized using a spinning-disk Olympus confocal fluorescence (Model IX81) motorized inverted research microscope.
Visualization of cell death in murine macrophages and human lung epithelial cells was conducted using ethidium-homodimer /Syto 10 stain after exposure to sodium molybdate (1, 3, 5 or 8 mM) or ce-MoS2 (up to 80 µg/mL) for 24 hours. Cells were washed once and incubated with 5 µM/ml Syto10/ and 1 µg/ml ethidium homodimer (Invitrogen) in phosphate buffered saline for 5 min (macrophages) or 15 min (lung epithelial cells). Images were visualized using a spinning-disk Olympus confocal fluorescence (Model IX81) motorized inverted research microscope.