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. 2007 Mar 21;81(11):5705–5713. doi: 10.1128/JVI.00076-07

FIG. 7.

FIG. 7.

Trastuzumab inhibits activation of PI3K/Akt kinase and mitogen-activated protein kinase kinase in MCF7-HA-BARF0 cells. (A) MCF7-Neor or MCF7-HA-BARF0 cells were cultured in the presence of 10 μg/ml trastuzumab, 20 μM U0126, or 20 μM LY294002 for 48 h. Cell lysates were separated by 12% SDS-PAGE and transferred onto nitrocellulose membrane for immunoblotting with phospho-ERK1/2, ERK1/2, phospho-Akt, Akt kinases, or HA antibodies. α-Tubulin was detected as an internal control. (B) MCF7-Neor or MCF7-HA-BARF0 cells (1 × 104 cells/well) were seeded in 6-cm dishes in culture medium containing 0.33% low-melting-temperature agarose over a 0.5% agarose layer and cultured in the presence of DMSO, trastuzumab (10 μg/ml), U0126 (20 μM), or LY294002(20 μM) for 2 weeks. Colonies were then stained with 0.05% crystal violet.