Multi-gene reporter array assay in co-cultures of reporter-construct-transfected FBs.

<p>The different FBs were either co-cultured with themselves or with NCI-H1437 or Calu-1 tumor cells. For each co-culture three independent experiments were performed. (A) Overview (heat map) on Luciferase signals of each of the 45 reporter constructs in transfected FBs (HDF, NF1 and CAF1). Reporters are indicated at the top. Culture conditions are depicted at the right side. Clustering method: UPGMA; distance measure: euclidean; ordering weight: average value. Red arrows indicate the six most significantly upregulated signaling pathways upon co-cultivation which are shown in detail in B. (B) Underlined FBs on the X-axis indicate the cell line which had been transfected with the corresponding reporter construct depicted in the header of each diagram. In all cultures either the same number of transfected FBs were co-cultured with the same number of non-transfected FBs (HDF+HDF, NF1+NF1 and CAF1+CAF1) or accordingly, with the same number of cells of non-transfected tumor cell line (HDF+Calu-1, HDF+H1437 etc.). Controls are depicted in green, Calu-1 co-cultures in purple and NCI-H1437 co-cultures in orange. Statistical analysis was performed by unpaired comparison of control samples (HDF+HDF, NF1+NF1, CAF1+CAF1) with respective co-culture samples by using Student’s t-test (*p<0.05, **p<0.01, ***p<0.001; n.s.: not significant). LI = luminous intensity.