Figure 2

Macrophage depletion correlates with reduced mesenchymal marker expression in F9-teratocarcinomas. (A) Relative gene expression (mRNA levels) of E-cadherin (cdh1), N-cadherin (cdh2), vimentin (vim), Snail and Twist in clodrolip treated vs. control tumors; n = 5-6, bars: means ± SEM. The tumors analyzed in this experiment were collected from two independent depletion experiments. Non-responders were excluded. (B) Relative gene expression (mRNA levels) of Csfr-1, E-cadherin (cdh1) and N-cadherin (cdh2) in size matched clodrolip treated vs. control tumors; n = 5-6, bars: means ± SEM. The tumors analyzed in this figure were collected from a single depletion experiment. The table shows individual volumes and gene expression levels of paired treated vs. control tumors. (C) Western blot analysis of E-cadherin, total β-catenin and active β-catenin expression in control and clodrolip treated tumors; n = 5. (D) Quantification of the western blot is shown in Figure C. (E) Immunofluorescence analysis of CD68⁺ macrophages (green), E-cadherin (red, left panel), β-catenin (red, mid panel) and fibronectin (red, right panel) in a macrophage poor area (upper panel, magnification 100×) and in a macrophage rich area (lower panel, magnification 100×) in a control tumor section (left picture, magnification 10×). The nuclei were stained with DAPI (blue). Scale bar = 0.02 mm. The graphs indicate correlations between CD68⁺ density and expression of the annotated EMT markers. Each data point represents analysis of one tumor section; n = 3-5/group. The linear regression coefficient R is indicated.