MTT Assay

Transfected PANC-1 cells were seeded in a 96-well plate at 5 × 10³ cells/well (180 μL/well). The plate was incubated overnight at 37°C in an atmosphere containing 5% CO₂ for the cells to adhere. The cells were then treated with GEM (0, 0.5, 1.5, or 15 μM) for 0, 24, 48, or 72 h. After GEM treatment, 20 μL of 5 mg/mL MTT solution (PAB180013, Bioswamp) was added to each well and the cells were further incubated for 4 h. Thereafter, the solution in the wells was aspirated and 150 μL of dimethyl sulfoxide was added to each well. The plate was gently shaken for 10 min and the absorbance of the wells was measured at 490 nm using a plate reader to evaluate relative cell proliferation.