Fig. 1

Cation channels are up-modulated in CAFs and involved in fibroblast activation. a Heatmap (bottom) reporting normalized enrichment scores (NES) for gene sets related to voltage gated channels and bar plot (top) reporting mean + sd, as calculated by GSEA on three independent datasets of prostate cancer patient-derived CAFs vs. matched NPFs. b qRT-PCR showing CACNA1H, CACNB1, CACNB3, SCN2A, SCN1B, and KCNS3 expression levels in an independent setting of three CAF and matched NPF cultures. Data were reported as relative expression compared to NPF and were representative of three independent experiments. c Western blotting analysis showing selected ion channel protein levels in a pair of matched CAFs and NPFs. β-actin was used as endogenous control. d qRT-PCR indicating relative expression levels of α-SMA, FAP and COL1A1 in NPF#1, NPF#2 and WPMY-1 fibroblasts exposed to CM of DU145 cells with respect to control fibroblasts. e Western blotting showing α-SMA, FAP and COL1A1 expression levels in WPMY-1 fibroblasts exposed or not to CM of DU145 cells. β-tubulin was used as endogenous control. f Immunofluorescence microphotographs showing α-SMA (green) and Col1a1 (red) expression in NPF and NPF exposed to CM of DU145 cells. Nuclei counterstained with DAPI (blue). Scale bar, 50 µm. g Cation channel mRNA expression levels in NPF#1, NPF#2 and WPMY-1 fibroblasts exposed to CM of DU145 cells with respect to untreated fibroblasts. h Western blotting displaying cation channel protein levels in NPF#1 and WPMY-1 fibroblasts exposed or not to CM of DU145 cells. β-tubulin was used as endogenous control. 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