Skip to main content
Figure 2 | Breast Cancer Research

Figure 2

From: Induction of epithelial to mesenchymal transition in PMC42-LA human breast carcinoma cells by carcinoma-associated fibroblast secreted factors

Figure 2

Effect of fibroblasts on major protein expression by PMC42-LA. Western blot analysis for a range of markers on control PMC42-LA organoids (control), and organoids cultured with NMFs beneath the filter (lane 1), with CAFs beneath the filter (lane 2), in medium conditioned by NMFs (lane 3), or in medium conditioned by CAFs (lane 4). Controls had no fibroblasts or fibroblast conditioned-medium. Immunocytochemistry was performed to view organization within organoid structures, and ethidium bromide was used to stain nuclei (red). (a) E-cadherin was not significantly upregulated in any of the fibroblast conditions, with no obvious changes in organization. (b) α-SMA expression was found to be significantly upregulated by all fibroblast conditions, with no obvious changes in organization. (c) Cytokeratin 14 expression was found to be significantly induced by all fibroblast conditions, with no obvious changes in organization. (d) Vimentin expression was also found to be significantly upregulated in all fibroblast conditions, with slight changes in organoid organization detected by immunocytochemistry. In control PMC42-LA cells (control lane in panel d), intermediate filament protein vimentin is localized in the cytoplasm of cells throughout organoids. With NMFs beneath the filter (panel d lane 1), vimentin remains localized in the cytoplasm of PMC42-LA cells and vimentin-positive cells are more organized, visible only on outer layer of organoids. With CAFs beneath the filter (panel d lane 2), the same organizational changes are observed. With NMF-conditioned media (panel d lane 3), the same organizational changes are observed. With CAF-conditioned medium (panel d lane 4), no change in organization is observed, with vimentin expression throughout organoids. CAF, cancer-associated fibroblast; NMF, normal mammary fibroblast; SMA, smooth muscle actin.

Back to article page