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. Author manuscript; available in PMC: 2013 Jun 11.
Published in final edited form as: Nat Med. 2012 Sep;18(9):1359–1368. doi: 10.1038/nm.2890

Figure 1.

Figure 1

Genotoxic damage to primary prostate fibroblasts induces expression of a spectrum of secreted proteins that includes WNT16B. (a) Schematic of the prostate cancer treatment regimen comprising a pretreatment prostate biopsy and four cycles of neoadjuvant DOC and MIT chemotherapy followed by radical prostatectomy. (b) DNA damage foci in human prostate tissues collected before and after chemotherapy. Tissue sections were probed with antibodies recognizing γ-H2AX (red and pink signals), and nuclei were counterstained with Hoechst 33342 (blue). Gl, gland lumen; e, epithelium; s, stroma. Scale bars, 50 µm. (c) Analysis of gene expression changes in prostate fibroblasts by transcript microarray quantification. The heatmap depicts the relative mRNA levels after exposure to H2O2, BLEO or RAD compared to vehicle-treated cells. Columns are replicate experiments. WNT16B is highlighted in bold for emphasis. (d) Measurements of WNT16B expression by qRT-PCR in prostate fibroblasts. Shown are the log2 transcript measurements before (Pre) or after exposure to the indicated factors relative to vehicle-treated control cells. Data are mean ± s.e.m. of triplicates. The P value was calculated by analysis of variance (ANOVA) followed by t test. (e) WNT16B protein expression in prostate PSC27 fibroblast extracellular conditioned medium (CM) or in cell lysates (IC) after genotoxic exposures. β-actin is a loading control. (f) Immunohistochemical analysis of WNT16B expression in prostate fibroblasts before (Pre) and after exposure to MIT or RAD. Brown chromogen indicates WNT16B expression. Scale bars, 50 µm. (g) Expression of Wnt family members in prostate fibroblasts after exposure to DNA-damaging agents. Transcript quantification was determined by microarray hybridization. Columns represent independent replicate experiments. WNT16B is listed in bold for emphasis. (h) WNT16B expression by qRT-PCR in PSC27 fibroblasts and prostate cancer cell lines after the indicated genotoxic exposure relative to pretreatment transcript amounts. Data are mean ± s.e.m.