Fig. 3.

Antiarrhythmics counteract the activated state of prostate CAFs. a Western blotting and relative quantification showing protein levels of fibroblast activation markers (α-SMA and COL1A1) in CAFs treated for 48 h with sub-toxic doses of antiarrhythmics. β-tubulin was used as endogenous control. b Bar plots showing the wound-healing rate assessed by scratch assay on CAFs exposed to antiarrhythmics. Data are reported as wound healing ratio at 24 h compared to 0 h point. c Representative immunofluorescence microphotographs (upper panel) showing the organization of β-actin cytoskeleton (green) and p-FAK (red) in CAFs treated with verapamil as compared to untreated. Scale bar, 50 μm. Western blotting analysis (lower panel) showing p-FAK, FAK protein levels in CAFs treated with sub-toxic doses of antiarrhythmics. β-tubulin was used as endogenous control. d Representative images (upper panel) showing 3D-collagen gel remodeling of CAFs exposed to sub-toxic doses of antiarrhythmics. NPFs were used as negative control. The dotted lines define gel areas. Bar plots (lower panel) showing ECM remodeling ratio of treated CAFs assessed by 3D-collagen gel assay. Data are reported as ECM remodeling ratio at 48 h compared to 0 h point. e Western blotting showing levels of pro-MMP2 and active-MMP2 in CM from CAFs exposed or not to sub-toxic doses of antiarrhythmics and, pro-MMP2 intracellular levels in treated cells. Gapdh was used as endogenous control for cell lysate. Results reported in the figure represent the mean (+ SD) of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.005, Student’s t-test