Figure 6

Morphologic and immunofluorescence characterization of EMT in MCF-7-MEK5 cells. (a) Morphologic comparison of MCF-7-VEC and MCF-7-MEK5 cells stained by Crystal violet assay. Representative colonies from clonogenicity assay were visualized after crystal violet staining for morphology. (b) MCF-7, MCF-7-MEK5, and MDA-MB231 cells were cultured in eight-well chamber slide for 48 hours. Indirect immunofluorescence was carried out to examine the expression of E-cadherin and F-actin, as described in the Materials and methods section. The nucleus was counter-stained with DAPI. Subpanels 1 through 4 are representative images of E-cadherin (1), F-actin (2), nucleus (3), and a merge of all three (4) in MCF-7-MEK5 (MEK5) cells. Subpanels 5 through 8 are representative images of E-cadherin (5), F-actin (6), nucleus (7), and a merge of all three (8) in MDA-MB231 (MDA) cells. Subpanels 9 to 12 were representative images of E-cadherin (9), F-actin (10), nucleus (11), and a merge of all three (12) in MCF-7 cells. Pseudocolors were assigned as follows: red, E-cadherin; green, vimentin; green, F-actin; and blue, nucleus. (c) MCF-7, MCF-7-MEK5, and MDA-MB231 cells were cultured in eight-well chamber slide for 48 hours. Indirect immunofluorescence was carried out to examine the expression of vimentin, as described in the Materials and methods section. The nucleus was counter-stained with DAPI. Subpanels 1 through 3 are representative images of vimentin (1), nucleus (2), and a merge of the two (3) in MCF-7-MEK5 (MEK5) cells. Subpanels 4 through 6 are representative images of vimentin (4), nucleus (5), and a merge of the two (6) in MDA-MB231 (MDA) cells. Panels 7 through 9 are representative images of vimentin (7), nucleus (8), and a merge of the two (9) in MCF-7 cells. Pseudocolors were assigned as follows: red, E-cadherin; green, vimentin; green, F-actin; and blue, nucleus. MEK, mitogen-activated protein kinase kinase.