Quantitative RT–PCR
FCS
CERVICAL CANCER
The human cervical immortalized squamous cell line Ect1/E6E7 and the cervical cancer cell line HeLa were recently acquired from ATCC and grown in DMEM containing 10% foetal calf serum (FCS, Lonza, Ambroise, France). Total RNA was extracted and purified by applying RNAsimple Total RNA Kit (Tiangen Biotech). For reverse transcription, the FastKing RT kit (with gDNase) (Tiangen Biotech) was used. qRT–PCR was carried out using SuperReal PreMix Plus (SYBR Green) from Tiangen Biotech according to the manufacturer's instructions. Each assay was duplicated at least three times for each sample. The 2