Effects of wortmannin on IL-6-induced phosphorylation of Akt (A) and IL-6-dependent growth of 7TD1 cells (B). 7TD1 cells were starved in IL-6-free medium for 6 h. Then the cells were pretreated with wortmannin or DMSO of equal volume for 30 min at 37°C before IL-6 was added into the medium. (A) The cells were cultured for 5 min in the presence of IL-6. After the cells were collected and washed, whole-cell extracts were prepared and subjected to Western-blotting assay. (B) The cells were seeded into 96-well plate (2 × 103 cells per well) and cultured in the presence of different dose of IL-6 for 72 h. Afterwards, MTT assay was performed to determine the effect of wortmannin on IL-6-dependent growth of 7TD1 cells. This figure is representative of three separate experiments.