Figure 1. EPO does not affect human breast cancer cell lines in vitro.
(A) Indicated cell lines were cultured in the presence of increasing concentrations of EPO (1, 5, and 10 IU/ml, replenished every other day), and proliferation was detected by MTT assay. (B) Cell lines were cultured in the presence of PBS or EPO (1 IU/ml) for 16 hours and EdU for 1 hour. They were then analyzed for EdU incorporation by flow cytometry. (C) Indicated cell lines were treated with vehicle, etoposide (50 μM), or etoposide (50 μM) and EPO (10 IU/ml) for 24 hours. Whole-cell extracts were Western blotted with the indicated antibodies. (D) Indicated cell lines were treated with DMSO, etoposide (50 μM), or etoposide (50 μM) and EPO (10 IU/ml) for 24 hours, then stained with PI and antibodies against annexin V. The percentages of cells that were annexin V positive and PI negative were quantified by flow cytometry. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.
