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. 2007 Aug;18(8):2838–2851. doi: 10.1091/mbc.E06-09-0875

Figure 5.

Figure 5.

5T4 antigen is localized at the plasma membrane in E-cadherin null ES cells. (a) D3, E-cadherin null (Ecad−/−) and N-cadherin null (Ncad−/−) ES cells were assessed for expression of cell surface 5T4 antigen in the presence of LIF (+LIF) and in the absence of LIF for 3 d (−LIF) in gelatin-treated plates using fluorescent flow cytometry analysis. 5T4 antigen (open population); isotype control (closed population). (b) (i) Immunofluorescence microscopy analysis of 5T4 antigen expression in E-cadherin null ES cells (DAPI shows total nuclei in the field of view). Bar, 10 μm. (ii) Enlarged image of the cells marked in b, pane i, demonstrating polarized expression of 5T4 in E-cadherin null ES cells. (c) Undifferentiated Ecad−/− ES cells were transfected with 2 μg of either (i) control (pCMV-neo) vector or (ii) vector containing full-length E-cadherin cDNA (pCMV-Ecad) using the Amaxa electroporation system and assessed for cellular phenotype by phase-contrast microscopy. (d) Cells transfected with (i) pCMV-neo vector or (ii) pCMV-Ecad were assessed for expression of cell surface E-cadherin (E-cad probe) and 5T4 antigen (5T4 probe) using fluorescent flow cytometry as described above. (e) Immunofluorescence microscopy analysis of 5T4 antigen expression in E-cadherin null ES cells transfected with pCMV-Ecad vector (DAPI shows total nuclei in the field of view). Bar, 5 μm.