Next Generation Sequencing Facilitates Quantitative Analysis of Control and beta-catenin Deficient Cancer-associated Fibroblasts Transcriptomes

D4M BrafV600E; Ptenlox5/lox5 melanoma cells mixed with uninduced control Fb (Ctnnb1+/+; Col1a2-CreER) or mutant bcat/Fb (Ctnnb1fl/fl; Col1a2-CreER) were injected intradermally into flanks of control Ctnnb1+/+; Col1a2-CreER or Ctnnb1 fl/fl; Col1a2-CreER mice, respectively. Tumors were allowed to grow to a volume of approximately 62.5 cubic millimeters when tamoxifen (TAM) was then administrated to induce ß-catenin ablation in mutant bcat/Fb CAFs. Control CAFs or bcat/CAFs were collected from BrafV600E; Ptenlox5/lox5 melanoma tumors using PDGFRa as a marker for fibroblasts. Normal fibroblasts were collected from B6 mouse skin. Overall design: mRNA profiles of normal fibroblasts; control CAF and bcat/CAF were generated by Illumina HiSeq-based next generation sequencing in triplicate.