Figure 4.

GKT137831 attenuates TGFβ1‐mediated fibroblast activation. Primary prostate human fibroblasts were treated with bFGF or TGFβ1 in the presence of 30 μM GKT137831 (GKT) or DMSO equivalent (control, ctrl) for 48 hr before (a) qPCR of Nox1 or Nox4 expression relative to the housekeeping gene TBP, (b) Western blotting of total cell extracts using the indicated antibodies and GAPDH as loading control, (c) determination of extracellular H₂O₂ levels via Amplex Red assay, (d) quantification of intracellular ROS via CM‐H₂DCFDA staining, (e) qPCR determination of the indicated CAF markers, and (f,g) analysis of migration using Boyden chamber transwell assays. (a,c–e,g) Data represent mean + SEM of at least four independent experiments using primary fibroblasts isolated from different donors. Statistical significance is shown (**p < 0.01; ***p < 0.001). (b) GAPDH served as loading control. (b,f) Images are representative of 3 independent experiments using cells isolated from different donors. (c,d) RFU, relative fluorescent units. Statistical significance is shown (*p < 0.05; **p < 0.01; ***p < 0.001).