Cell Proliferation Assay

A Cell Counting Kit-8 (CCK8) assay was used to detect cellular proliferation. The transfected cells were seeded into 96-well plates at a density of 5,000 cells per well in triplicate wells. Cell viability was measured using the CCK8 system (GIBCO) at 0, 24, 48, 72, and 96 h after seeding, according to the manufacturer’s instructions.

For the colony formation assay, transfected cells were seeded into six-well plates at a density of 2,000 cells per well and maintained in DMEM containing 10% FBS for 10 days. The colonies were imaged and counted after they were fixed and stained.