Fig. 4

Immunofluorescence (IF) validation of cytokine data in tumor and stroma tissue. a, b Representative IF images of nerve growth factor beta (β-NGF) or IL-17A receptor (IL-17A-R) (magenta) counterstained with 4',6-diamidino-2-phenylindole (DAPI) (green); ×20 objective; scale bar = 100 um. Composite is an enlarged image of the area demarked by the yellow window; scale bar = 15 um. c Quantitation of β-NGF normalized to the total number of cells. β-NGF is elevated in the stroma of PyMT/Col1a1 tumors. Celecoxib (Clxb) decreases β-NGF in both the epithelium and stroma in PyMT/Col1a1 tumors. β-NGF is higher in the tumor epithelium, consistent with its known role in promoting survival and proliferation of epithelial breast cancer cells. d Quantitation of IL-17A-R normalized to the total number of cells. IL-17AR is elevated in the stroma of PyMT/Col1a1 tumors, consistent with its role in macrophage/neutrophil recruitment. Celecoxib treatment decreases IL-17A-R only in the stroma of PyMT/Col1a1 tumors. Raw data are depicted; *p < 0.05, ***p < 0.001, ****p < 0.0001; n = 5 mice per arm; at least 8 image fields analyzed per 2–3 tumors per animal; mixed linear model. Veh vehicle