Fig 1: Immunohistochemical analysis of Adam12 protein expression in tissue microarrays.(A) Adam12 protein expression is undetectable in the granule cell layer of the brain (negative control tissue). (B) Stromal fibroblasts (arrows) surrounding normal pancreatic duct do not label Adam12. (C) Cancer associated fibroblasts (arrows) in a primary pancreatic adenocarcinoma are strongly positive for Adam12 protein; magnification, 20×.
Fig 2: Bisulfite sequencing analysis of ADAM12.(A) Top: ADAM12 gene structure and distribution of CpG dinucleotides. Short vertical bars represent CpG sites. Arrow points to transcriptional start site. Below: Bisulfite genomic sequencing analysis in pancreatic CAFs and control fibroblasts. Open circles represent unmethylated CpG sites, solid black circles methylated CpG sites, and hatched circles partially methylated CpG sites. (B) Bisulfite sequencing chromatograms of the ADAM12 promoter in a pancreatic CAF (CAF19) and control fibroblast line (HPNE). Arrows point to cytosine residues.
Fig 3: Analysis of ADAM12 mRNA expression in pancreatic CAFs and control fibroblasts.(A) Affymetrix exon array analysis of ADAM12 mRNA expression in pancreatic CAFs and control fibroblasts. (B) Quantitative RT-PCR analysis of ADAM12 mRNA expression in pancreatic CAFs and control fibroblasts after normalization to GAPDH levels. Each assay was performed in triplicate. Data are means of three independent experiments; bars are SD values.
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