Cell growth determination kit

MTT assay was preformed using the Cell Growth Determination Kit (Sigma) as previously described 9 (link). Briefly, cells were seeded onto 96-well plates at 1x10³ cells/well. After 24 hours of culture, cells were transfected with shRNA-LncRNA NONHSAT141924 or pcDNA-LncRNA NONHSAT141924 and treated with paclitaxel in 100 µL media. After 48 hours of treatment, 10 µL MTT (methylthiazol tetrazolium) solution was added into the media. After incubation at 37°C for 4 hours, the supernatants were removed, and cells were incubated with 100 µL of MTT solvent. Absorbance at 570 nm was spectrophotometrically measured using a BioTek ELx800. The assays were performed independently and repeated at least three times.

Cell viability was determined with the commercial, MTT-based, Cell Growth Determination Kit, (Sigma-Aldrich, St Louis, MO, USA) according to the product protocol. Briefly, cells were cultured in 96-well plates at a density of 5 × 10³ cells in the normal culture medium (100 μL) for 24 h after which the medium (75 μL) was changed and the cells were exposed to 0.2, 1, 5, 10, and 25 mg/mL of bilberry added in 25 µL of PBS. Phosphate-buffered saline (PBS 25 µL/100 µL) was used as a control. After 24 or 72 h of incubation, MTT solution was added and incubation continued for 3–4 h. Culture media was removed, and the cell cultures washed twice with PBS before adding isopropanol (MTT solvent) to avoid the effects of anthocyanin pigments on the measurements with an ELISA reader at 550 nm. There were two (HSC-3 cells) or three (HMK and IHGK cells) independent experiments with six replicates in each assay.

Cell viability was measured using an MTT-based Cell Growth Determination kit (# GDC1; Sigma, St Louis MO, USA). The MTT solution was added to each well in sterile conditions (the final concentration was 10% of total volume) and the plates were incubated for 4 h at 37 °C. The formazan crystals formed and were dissolved in solubilization solution (1:1); the purple formazan crystals were formed from yellow MTT by succinate dehydrogenase in viable cells. The absorbance of the dissolved formazan product was measured at a 570 nm background corrected to 690 nm using a microplate reader. Each experiment was performed in triplicate and statistically significant differences were evaluated using a Student t-test.

Cell viability was determined with the Cell Growth Determination kit (Sigma-Aldrich, St. Louis, MO, USA) following the manufacturer’s instructions. The experiments were carried out on 96-well plates (Corning, NY, USA), 40,000 cells were seeded per well and infected with DENV2 at MOI of 5. After different hours post-infection, we used 50 μg of 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) per sample. The MTT assay involves the conversion of the water-soluble yellow dye MTT to an insoluble purple formazan by the action of mitochondrial reductase. Formazan is then solubilized in isopropanol and the concentration determined by optical density at 570 nm in an ELISA-type plate reader (IMarkTM, Bio-Rad, Hercules, CA, USA).

Cell viability in the presence or absence of different inhibitors or dimethyl sulfoxide (DMSO) was measured by a 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide (MTT)-based assay (Cell Growth Determination Kit, Sigma–Aldrich) in accordance with the manufacturer's protocol. CEF or DF-1 cells were seeded into 96-well plates and pretreated with different inhibitors or DMSO for various lengths of time. After each incubation period, 10 μL of MTT labeling mixture was added to each well, and cells were incubated at 37°C for 4 h. Absorbance was measured by an enzyme-linked immunosorbent assay reader at a wavelength of 490 nm. Data represent the average of at least three independent experiments.