Figure 3.

Sensitisation of human ovarian carcinoma cells to CB1954 by NfsA or NfsB. (A) SKOV3 cells were incubated in a medium containing 50 μMCB1954, 200 μM NADH or NADPH and a range of concentrations of purified NfsA or NfsB proteins. The components were mixed at the start of a 4-h incubation period at 37°C, after which the medium was changed. Cell viability was determined 2 days later using the MTT assay. One representative experiment of three is shown. (B) To confirm the enzyme concentrations used in panel A, 200 and 50 ng of the purified proteins were analysed by SDS-PAGE and stained with Coomassie blue. (C) SKOV3 cells were infected using 0, 30, 100 or 300 p.f.u. per cell of adenovirus vectors CTL102 or AdSV042, expressing NfsB or NfsA, respectively. After 2 days, they were exposed to a range of CB1954 concentrations for 4 h, before determination of cell viability using MTT assay 2 days later. (The values used are representative of at least three experiments.)