Figure 3

GAG-hed causes repression of HPV18 E6/E7 transcription in HeLa cells. (A) Northern blot was performed with total RNA obtained from HeLa cells treated with 2 mg/ml of GAG-hed and harvested 72 or 96 h later. During treatment, cells were grown in media with 2% serum. Control cells were harvested after 72 h. Gel image showing similar RNA loading in gel was taken before transfer and shown as reference. (B) Cells were treated with the indicated increasing concentrations of GAG-hed and total RNA was extracted after 48 h. RT-PCR assays were performed and DNA products were separated on agarose gels. RNA used as negative control was from C33-A cell line. E6/E7 transcript is denoted as a 270 bp band (top); same mRNAs were used to test the expression of β2-microglobulin, denoted as a 100 bp band (bottom). (C) Plot represents densitometryc data normalized against microglobulin optical density.