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. 2015 Feb 25;4(2):e978709. doi: 10.4161/2162402X.2014.978709

Figure 5.

Figure 5

(See previous page). Matrix architecture of GU and skin tumor stroma is characterized by distinct patterns of collagen deposition. Genitourinary (GU) and cutaneous tumors from RETAAD mice were examined for differences in stromal and matrix composition by immunostaining and fluorescence cytometry (A-C) and high-resolution 2-photon microscopy (D-E). (A) Representative dot plots showing flow cytometry gating strategy for CD45− CD31+ endothelial cells and CD45− CD31− PDGFRα+ tumor-associated fibroblasts (TAFs). Flow cytometry analysis was performed to quantify (B) CD45− CD31+ endothelial cells and (C) CD45− CD31− PDGFRα+ TAFs present in GU and skin tumors. (D) High-resolution images of collagen fibres within the GU and skin tumor stroma were generated by second harmonic generation (SHG) using a 2-photon microscope (E) Immunofluorescence imaging were performed to examine T cells interactions with the GU tumor stroma. Data from panels: (B and C) are pooled from 2 separate experiments with 4-5 mice per group in each experiment (n = 9-15/group). Bars represent mean ± SD. Statistical analyses were performed using the unpaired two-tailed Student's t-test; **p<0.01. Images from (D and E) are representative of 5 independent experiments (n = 5/group). Scale bars in (D) and (E) represent 75 μm and 50 μm, respectively.