Jc 10 fluorescent probe

Mitochondrial membrane potential was assayed in THP-1 and A549 cells using the JC-10 fluorescent probe (Sigma). Cells were seeded into 96-well culture plates at a density of 3.10⁴ (A549) or 10⁵ (THP-1) cells/well. PMA (10 ng/mL) was added to culture medium of THP-1 cells to induce their differentiation into macrophages. The following day, all cells were incubated with 100 µg/mL NPs for 4 h. Then, the cell culture supernatant was removed and the JC-10 probe (100 µL) was added to the cells for 1 h. Fluorescence of the samples was then measured (functional mitochondria: λ_ex = 540 nm, λ_em = 590 nm; non functional mitochondria: λ_ex = 490 nm, λ_em = 525 nm), and the ratio of fluorescence intensity at 525 nm to fluorescence intensity at 590 nm was calculated for each sample. Then, data were expressed as the percentage of fluorescence of NP-exposed cells relative to the fluorescence ratio of non-exposed control cells.

Mitochondrial membrane potential was assayed using the JC-10 fluorescent probe (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer’s instructions. Cells were seeded into 96-well culture plates at a density of 10⁵ cells/well, differentiated into macrophages, and incubated with increasing concentrations of CDs for 4 h. After the CD treatment, the cell culture supernatant was removed and the JC-10 probe (100 μL) was added to the cells for 1 h. Then, fluorescence of the samples was measured (functional mitochondria: λ_ex = 540 nm, λ_em = 590 nm; non-functional mitochondria: λ_ex = 490 nm, λ_em = 525 nm). The ratio of fluorescence intensity at 525 nm to fluorescence intensity at 590 nm was calculated for each sample. Data were expressed as the fold change of CD-exposed cells relative to the non-exposed control cells.