Figure 3.
QrtPCR validation of selected mRNAs in CAF and NPF patient-matched samples and in fetal prostate. QrtPCR for candidate mRNAs was performed on the CAF and NPF samples used for Tag profiling (labelled CAF1), as well as six further matched pairs of functionally tested CAFs and NPFs (labelled 2–7). In all candidates, the Tag count data matched the QrtPCR transcript level and confirmed enrichment or depletion of candidate transcripts in the CAF/NPF sample used for Tag profiling. Of the CAF-enriched molecules: ASPN was increased in two and decreased in three CAF/NPF pairs, CTSK was increased in three CAF/NPF pairs, FBLN1 was increased in three and decreased in two CAF/NPF pairs, FN was increased in five CAF/NPF pairs, OGN was increased in five and decreased in one CAF/NPF pairs, PARVA was increased in two and decreased in four CAF/NPF pairs, ZEB1 was increased in one and decreased in two CAF/NPF pairs. Of the CAF-depleted molecules: CAV1 was decreased in four CAF/NPF pairs, CFH was increased in three and decreased in four CAF/NPF pairs, NKTR was increased in three and decreased in three CAF/NPF pairs, S100A6 was increased in two and decreased in five CAF/NPF pairs, STC1 was increased in one and decreased in four CAF/NPF pairs. As controls, CXCL12 was increased in five and decreased in two CAF/NPF pairs, while WDFC1 was decreased in seven CAF/NPF pairs (Supplementary Figure 1; Supplementary Table 1). In addition, transcript levels were measured in fetal human prostate (EMB) to determine relative mRNA levels, and there was good (but not complete) correlation with candidate Tag counts in the EMB library (Table 2).