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. 2020 Jan 28;9:e50740. doi: 10.7554/eLife.50740

Figure 1. Fibronectin expression is restricted to stromal cells in PyMT tumors, but not in tumors that have undergone EMT.

Figure 1.

In the MMTV-PyMT model, fibronectin is only expressed by stromal cells, if the tumor is (A) spontaneous (SP) or (B) results from an orthotopical implant (TP) of cells dissociated from a SP tumor. Fibronectin may also be expressed by all tumor cells after EMT has taken place in (C) the TC1 cell line, here transplanted subcutaneously, or in (D) a spontaneous melanoma, TiRP. Tumor pieces were fixed overnight with periodate-lysine-paraformaldehyde at 4°C. Then, tumor samples were embedded in 5% low-gelling temperature agarose prepared in phosphate-buffered saline (PBS). 350 μm slices were cut with a vibratome in a bath of ice-cold PBS. Immunostaining of surface markers was performed at 37°C for 15 min with antibodies specific to EpCAM-BV421, CD31-Biot (both BD Pharmingen) or Fibronectin (Abcam). CD31-Biot was revealed with a streptavidin-PE (BD Pharmingen) and fibronectin with a goat anti-rabbit-FITC (Invitrogen).