Fig. 1

EnAd genome replication in a multi-indication human cell line panel. Duplicate cultures of 23 test tumour cell lines, plus positive (A549, small cell lung carcinoma) and negative (CT26, mouse colon carcinoma) control cell lines were inoculated with 1ppc of EnAd or assay media alone (uninfected control) and cultured at 37 °C, 5% CO₂ for 3, 4, 8 or 11 days. At each time point, supernatants and cell lysates were harvested and frozen, before DNA extraction. qPCR was then run (triplicates) using the E3 primer/probe set. Data was background subtracted: mean genome quantity for each uninfected control triplicate was subtracted from the corresponding individual EnAd values. Genome quantity per cell was then calculated, and the mean of the two EnAd qPCR triplicates determined. Lysate and supernatant results were then combined to give a total genome detection value. The mean of the duplicate values at the time point showing maximal expression was plotted in the graph, with the SD represented by error bars. For the positive control cell line (A549), mean and SD was calculated across all runs at day 4 (n = 10 qPCR triplicates). For the negative control cell line (CT26), mean and SD was calculated across all runs at day 11 (n = 9 qPCR triplicates). Maximal genome expression was at day 8 for the following cell lines: HCT-116, SW620, SW480, PC-3, CFPAC-1, Caov-3, HT-1376, and at day 11 for the following cell lines: HT-29, DU145, LNCaP, Panc-1, Capan-2, BxPC-3, MDA-MB-231, MDA-MB-453, BT-20, MCF-7, SKOV3, OVCAR-3, PA-1, RT4, T24, UM-UC-3. Viral genome was undetectable at any time point with PA-1 cells