2.2. MTT Assays

The MTT assay was performed to detect the cytotoxicity of azacitidine. Briefly, 2 × 10⁴ cells were seeded into 96-well plates, incubated overnight, and exposed to azacitidine (0, 1, and 10 μM) for 1 or 24 h and harvested on the 72nd hour. MTT solution (0.5 mg/mL) was added to the wells and incubated for 3 h at 37 °C. The reaction was ended by the removal of the supernatant, followed by dissolving the formazan product in DMSO. Optical density was measured with an ELISA reader using a 570 nm filter. Each experiment was performed in triplicate and the cell viability of each group was calculated and analyzed compared to the control.