Figure 7.

Human breast cancer-associated fibroblasts and normal-associated fibroblasts have distinct inflammatory responses to soluble TGFBR3. (A) Conditioned media was collected after 48 h after either no treatment or treatment with 200 ng/mL of soluble TβRIII (sTβRIII). Two hundred microliters of conditioned medium was used to analyze protein expression of 105 human cytokines on R&D Systems Human Cytokine Array XL detection kit. (B) Quantification of pixel intensity by IR secondary scanning revealed distinct pixel density in select cytokines with a specific response to sTβRIII treatment of either NAFs or CAFs. Column graph represents mean of pixel density value replicates with SD error bars. (C) qPCR for selected cytokines demonstrates relative mRNA expression (2^−ΔΔCt) in cytokines from NAFs and CAFs treated with sTβRIII. mRNA is normalized to GAPDH levels and relative to control. The fold changes are given in log² scale. Column graph represents mean of three replicates with SD error bars. No statistical significance was observed in qPCR analysis. Gene abbreviations are defined as follow: CD40 (cluster of differentiation), SHBG (sex hormone binding globulin), GM-CSF (granulocyte macrophage colony-stimulating factor), IL-3 (interleukin 3).