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. 2019 Oct 4;55(6):1275–1286. doi: 10.3892/ijo.2019.4892

Figure 2.

Figure 2

Generation of MCF7-B2 cells. Infected cells with viral vectors expressing GFP (lanes 2 and 3), ephrin-B2-WT (B2-WT) (lanes 4 and 5) and ephrin-B2-5F (B2-5F) (lanes 7 and 8) were treated with two multiplicities of infection (MOI): MOI 25 (lanes 2, 4 and 7) and MOI 1.5 (lanes 3, 5 and 8). Total lysate from SKBR3 cells infected with human ephrin-B2 (EX-M0409-Lv105, Genecopoeia) (~50 kDa) was used as a positive control (C+). Non-infected MCF7 cells (N.I) were used as a negative control. GAPDH was used as a loading control. Ephrin-B2 detected with the anti-pan (A) ephrin B antibody or (B) anti-GFP antibody and visualized as an 80 kDa GFP-fusion protein indicated by arrowheads. Ephrin-B2 activation was verified by (C) immunoprecipitation and column charts, from this representative experiment, and representative immunoblots are shown. Ephrin-B2 expression was visualized by confocal microscopy at ×600 magnification. (D) Cell nuclei are indicated with DAPI staining. Scale bars, 30 µm.