Figure 3

Invasion of fibroblasts and cancer cells in 3D culturing conditions. (A) Phase contrast images of HMF3s fibroblasts grown in Matrigel in mono-culture. Notice sparse outgrowth of HMF3s cells. Scale bar = 300 μm. (B) Phase contrast images of MCF7S1 tumour cells grown in Matrigel in mono-culture. Notice absence of invasion in MCF7S1 cancer cells. Scale bar = 500 μm. (C) Invasion of MCF7S1 cancer cells in response to treatment with fibroblast CM. Notice pronounced invasion of cancer cells. Scale bar = 500 μm. (D) Mutual invasion of HMF3s fibroblasts [F] and MCF7S1 cancer cells [C] co-cultured in 3D Matrigel. Inset shows details of the invasive front. Scale bars = 500 and 100 μm, respectively. (E) Immunohistochemical detection of S100A4 in the invasive front of HMF3s fibroblasts grown in co-culture with MCF7S1 cells. Arrows point to invading cells. Scale bar = 100 μm. (F) Western blot analysis of cell lysates from 3D cultures showing increased S100A4 expression co-cultures. α-tubulin was used as a loading control. Note that the expected amount of α-tubulin in lane 3 is equal to the sum of lane 1 and 2. (G) Quantification of the blot shown in Figure 3F with tubulin adjusted to the cell amounts in each lane. The co-culture (3) represents 100%.