Colony-forming unit (CFU) Assay.

HeLa cells were seeded in triplicates with 2 × 10⁴ cells into a 24-well and siRNA-transfected in antibiotic-free medium. The experiment was performed 48 to 72 h posttransfection. After the infection, cells were washed with PBS and cultured with Gentamycin-containing medium. At 1, 6, or 8 h pi, cells were lysed in 0.1% (vol/vol) Triton X-100 in PBS and serial diluted in PBS. Diluted lysates were plated in technical duplicates on Agar plates and incubated for colony formation overnight at 37 °C. The number of colonies (within the linear range of the assay) was enumerated using a colony counter apparatus and software (aCOLyte 3, Synbiosis).