Celltiter glo luminescent

PMBL cell lines were treated for 24h with Doxo (0,1ng/ml), VC (1μg/ml) and RmAB (1μg/ml). According to manufacture's protocol the cell viability analysis was performed using the CellTiter-Glo® Luminescent (Promega, Mannheim, Germany.
T-tests were used for statistical comparisons and the statistical significance was defined as p <0.05.

The number of viable cells was assessed by measuring intracellular ATP to identify metabolically active cells, using the CellTiter-Glo luminescent cell viability assay (Promega). The number of viable cells after 5 days was assessed by measuring intracellular WST-8 formazan using the Cell Counting Kit-8 (Dojindo Laboratories, Kumamoto, Japan).

To measure ATP consumption, a CellTiter-Glo® luminescent cell viability assay (G7570, Promega) was used. Cells were seeded on 96-well plates (1000 cells/well, triplicates) and treated with specified conditions. After 5-day treatment, cell viability was examined according to the manufacturer’s instructions. In brief, the CellTiter-Glo® solution was applied to each well and incubated at room temperature for 10 min. The intensity of luminescence was measured using a microplate reader (EnSpire, PerkinElemer). Cell viability was calculated relative to DMSO treatment (100%). For the MTT proliferation assay, cells were seeded on 96-well plates (5000 cells/well, triplicates) and cultured under indicated conditions. For each condition, the reagent 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma-Aldrich) was added to the cells and incubated for 4 h at 37 °C (5% CO₂). The supernatants were discarded and replaced by 200 µL DMSO to dissolve the formazan product. Absorbance was measured at 570 nm in a plate reader (BMG Labtech).

All viability assays were run in 96-wellpates at 72 hours. IC50 viability assays were determined by Cell-Titer Blue (Promega catalog # G8080) and Cell Titer-Glo Luminescent (Promega catalog # G7570). EC50 viability assay was evaluated using the MTT Cell Proliferation Assay Kit (ATCC catalog # 30–1010K).

The inhibitory effect of different drugs was assessed following manufacturer’s protocol on i) NSCLC cell growth by measuring 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide inner salt (MTT, Sigma-Aldrich) dye absorbance of cells, and ii) LCSC cell growth by quantitation of the ATP present in metabolically active cells using CellTiter-Glo® Luminescent (Promega, Southampton, UK). Data were analyzed by the Chou-Talaly method (CalcuSyn software, Biosoft, Cambridge) to determine the combination index (CI), a well-established index of the interaction between two drugs. CI values of <1, =1, and >1 indicate synergistic, additive, and antagonistic effects, respectively.

The biocompatibility of modified pSiNPs on the hCMEC/D3 cell line was determined using a CellTiter-Glo^® luminescent cell viability assay (Promega, Alexandria, Australia). Briefly, hCMEC/D3 cells were seeded onto a 96-well white opaque polystyrene microplate (Sigma-Aldrich, Macquarie Park, Australia) at a density of 10,000 cells per well and maintained in the cell culture medium for 1 day. Subsequently, cells were treated with different concentrations (5, 10, 25, 50 µg/mL) of surface-modified pSiNPs. Cells without any treatment were used as control, and each condition was triplicated. After incubating cells for 48 h, a CellTiter-Glo^® luminescent cell viability assay was used to evaluate the cell viability. Briefly, 100 µL of the solution of the assay kit was added to 100 µL of the medium in each well, and the plates were gently shaken at RT for 15 min. Finally, the luminescence intensity of each sample was obtained using a PerkinElmer EnSpire multimode plate reader, and data were expressed as the mean and standard deviation of 3 replicates.

PMs were seeded in a 96-well plate (1 × 10⁴ cells/well), and A419259 (0.01, 0.1, 0.25, 0.5, and 1 μM) was added to the medium after 12 h. After 4 h, 100 μl of the CellTiter-Glo^® luminescent (G7570, Promega, USA) solution was added to the 96-well plate and mixed gently on an orbital shaker at room temperature for 10 min. The resulting fluorescence was measured using a multiscan spectrophotometer (TECAN, SPARK 10M).