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. 2014 Jan 2;124(2):553–563. doi: 10.1172/JCI69804

Figure 2. EPO decreases the percentage of breast cancer GEMMs living over time yet does not affect breast cancer GEMM cell lines in vitro.

Figure 2

(A) Kaplan-Meier survival curves of MMTV-Neu and C3-Tag mice randomized to saline or EPO (500 IU/kg BIW) injections. (MMTV-Neu: P = 0.05, C3-Tag: P = 0.04). Tumors were sectioned and H&E stained. (B) Indicated cell lines were cultured in the presence of increasing concentrations of EPO (1, 5, and 10 IU/ml, replenished to the culture every other day). and proliferation was detected by MTT assay. (C) 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. (D) 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. (E) Indicated cell lines were treated with DMSO, etoposide (50 μM), or etoposide (50 μM) and EPO (10 IU/ml) for 24 hours. Percentage of apoptotic cells was then determined by flow analysis using Alexa Fluor 488 annexin V/PI Cell Apoptosis Kit (Invitrogen). ***P ≤ 0.001.