96 well flat bottom culture plates

HaCaT cells (1 × 10⁴ cells/well) in 96-well flat-bottom culture plates (SPL Life Sciences, Pocheon, Korea) were treated with indicated doses of FSCP for 24 h with or without CoCl₂. Cell viability was determined using the colorimetric WST-1 conversion assay (EZ-Cytox assay kit, Daeil Lab Service, Seoul, Korea). WST-1 reagent (10 μL) was added to each well, after which cells were incubated for 2 h in a humidified incubator at 37°C under 5% CO₂. Absorbance of the formazan dye, generated by the reaction between dehydrogenase and WST-1 in metabolically active cells, was measured using a microplate reader (Tecan, Männedorf, Switzerland) at 450 nm according to the manufacturer's instructions. Percent cell viability was calculated. Experiments were performed at least thrice.

The TECs were seeded at a density of 8 × 10³ cells per well on 96-well flat-bottom culture plates (SPL Life Sciences, Pocheon, Republic of Korea). The cells were then treated with the specified doses of FCPs for a duration of 48 h, either in the presence or absence of Bay 11-7082 or Bay 11-7085 for 15 or 30 min. The assessment of cell proliferation was conducted using the colorimetric WST-1 conversion assay, namely the EZ-Cytox assay kit provided by Daeil Lab Service in Seoul, Republic of Korea. In each well, a 10 μL volume of WST-1 reagent was introduced, followed by incubation of the cells for a duration of 2 h. This incubation was carried out in a humidified incubator at a temperature of 37 °C, with a CO₂ concentration of 5%. The measurement of the absorbance of the formazan dye, which is produced through the reaction between dehydrogenase and WST-1 in metabolically active cells, was conducted using a microplate reader (Tecan, Männedorf, Switzerland) at a wavelength of 450 nm, following the guidelines provided by the manufacturer. Subsequently, the percentage of cell proliferation was determined through calculations. The experiments were conducted in triplicate.

After TECs (8 × 10³ cells/well) in 96-well flat-bottom culture plates (SPL Life Sciences, Pocheon, Korea) were treated with the indicated doses of FCP for 24 h with or without cisplatin. The cell viability was determined using the colorimetric WST-1 conversion assay (EZ-Cytox assay kit, Daeil Lab Service, Seoul, Korea). A WST-1 reagent (total 10 μL) was added to each well, and cells were incubated for 2 h in a humidified incubator at 37 °C under 5% CO₂. The absorbance of the formazan dye, generated by the reaction of dehydrogenase with WST-1 in the metabolically active cells, was measured using a microplate reader (Tecan, Männedorf, Switzerland) at 450 nm according to the manufacturer’s instructions, and the percent cell viability was calculated. The experiments were performed in triplicate.