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Fig. 3 | Breast Cancer Research

Fig. 3

From: Evidence of two distinct functionally specialized fibroblast lineages in breast stroma

Fig. 3

CD105high fibroblasts exhibit a transforming growth factor (TGF)β profile and mesenchymal stem-like properties. a Heat map representation of microarray analysis of 44 selected, differentially expressed genes in CD26high and CD105high fibroblasts (passage 9). Color key indicates centered and row-scaled normalized intensity values. b Overlap between the top 302 median ranked significantly and differentially expressed genes and previously published profiles of breast normal and tumor stroma, respectively. Bars indicate the number of overlapping genes in CD26high or CD105high cells compared to tumor or normal stroma, respectively. The overlap between genes expressed by CD105high cells and tumor stroma was statistically significant on analysis by Fisher’s exact test (p < 0.001). c Oil Red O staining of lipid droplets at day 15 of adipocyte differentiation in CD26high and CD105high cells, respectively, with nuclei stained with hematoxylin. d RT-qPCR of osteoblast and adipocyte marker gene expression (CCAAT/Enhancer binding protein, alpha (CEBPA), Collagen type 1 alpha 1 (Col1a1), and Runt-Related Transcription Factor 2 (Runx2)) in CD26high (light bars) and CD105high (dark bars) cells at day 3 of differentiation in passage 11 presented as gene expression relative to the geometric mean of two reference genes (UBC and B2m). The difference was statistically significant for CEBPA and Col1a1 in three different biopsies on analysis by unpaired Student’s t test at p < 0.05. e Alizarin Red staining and quantification of the mineralized matrix at day 15 after exposure to osteogenic induction medium (OIM, +) or control conditions without inducing factors (−). On analysis by unpaired Student’s t test the difference in Alizarin Red staining in samples representing four biopsies, two in passage 11 and two in passage 13, was not statistically significant in CD26high with and without osteogenic induction, but was significant at p < 0.05 in CD105high with and without induction (scale bar = 100 μm). Error bars represent mean +/− SD

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