Figure 4

TARS is secreted by SK - OV - 3 ovarian cancer cells. (A) SK-OV-3 cells were treated with TNF-α (50 ng/ml) or exposed to 2% O₂ for 24 h where indicated. Media was concentrated 20-fold to accommodate 25% onto the gel and compared to 5% of the cell lysate. Shown is a representative Western blot probed for TARS and β-tubulin, n = 4. (B) Cells were treated as in (A). After 24 h the level of TARS in the supernatant was determined by ELISA, *p < 0.05, n = 3. Cell membrane integrity was confirmed using the lactate dehydrogenase assay CytoTox-ONE™ and by lack of β-tubulin in media samples. (C) Relative mRNA levels for TARS, VEGF and IL-1β were determined by RT-qPCR following treatment as in (A); *p < 0.001, **p < 0.0001, n = 4.