Journal: Cancer cell

Article Title: Activation of translation complex eIF4F is essential for the genesis and maintenance of the malignant phenotype in human mammary epithelial cells.

doi: 10.1016/j.ccr.2004.05.024

Figure Lengend Snippet: Figure 3. eIF4E confers HMEC with the ability to undergo anchorage-dependent and -indepen- dent clonal proliferation

Article Snippet: To identify the HA tag by immunohistochemical staining, 4 m sections were deparaffinized, boiledImmunoblot analysis with antigen unmasking solution for 10 min, incubated with anti-HA antibodyTo analyze the components of eIF4F, cells were lysed by three freeze-thaw prelabeled with Alexa 488 fluorophore (Molecular Probes, 1:50) for 2 hr, andcycles, and equal amounts of cell extract protein per lane were subjected counterstained for 20 min with DAPI (Sigma).to SDS-PAGE, blotted onto nitrocellulose membranes, and identified using antibodies directed against eIF4E (mouse monoclonal antibody, 1:500, StatisticsTransduction Laboratories), phospho-elF4E (rabbit polyclonal antibody, Statistical analyses were performed using one-way analysis of variance1:1000, Cell Signaling), and eIF4G1 (rabbit polyclonal antibody, 1:4000, de(ANOVA) with Dunnett’s multiple comparison test (S-PLUS Guide to Statisti-veloped in the laboratory of N.S.).

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