Cell Viability Assay

HT-29 over-expressing or inhibiting of the miR-378a-3p and their respective cell line controls were seeded (3 × 10⁵ cells) in 6-wells plates with DMEM, FBS (10%) and Penicillin/Streptomycin (1%). Afterward, cells were trypsinized at 6, 12, 24, 48, and 72 h and the number of viable cells (unstained) were counted by trypan blue stain (Thermofisher Scientific Carlsbad, CA) using a Neubauer Camera. Additionally, viable cells were determined by flow cytometry (FACSCanto Becton Dickinson Biosciences, San Jose CA, USA) using propidium iodide (PI) stain (10 μg/mL) (ThermoFisher Carlsbad, CA, USA) at 24, 48, and 72 h in four duplicate independent experiments.