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Fibroblast-to-myofibroblast -differentiation was induced by 1 ng/ml TGFβ1, whereas control cells were incubated with 1 ng/ml basic fibroblast growth factor (b FGF) in RPMI 1640 (PAA Laboratories) supplemented with 1% charcoal-treated bovine calf serum (Hy Clone Laboratories, Logan, UT) and antibiotics for 72 h as described (7).

For -differentiated Pr SC were transduced with lentiviral particles at multiplicity of infection 8 and analyzed 6 d after transfection (media were replaced by fresh medium containing b FGF or TGFβ1 after 72 h).

Human prostatic organoids were established from radical prostatectomy from tissue-wedges from the ventral prostate showing no histological signs of malignancy.

Human primary Pr SC were established as described previously (23) and cultured in stromal cell growth medium (Quantum 333; PAA Laboratories, Pasching, Austria).Furthermore, Rho A has been shown to modulate TGFβ-induced smooth muscle cell differentiation via cross talk with Smad (19).The PDE5 inhibitors vardenafil and sildenafil have been shown to inhibit Rho A/ROCK in a NO/c GMP-dependent protein kinase G (PKG)-dependent manner in the bladder stroma and in vascular smooth muscle cells, respectively (20, 21).We and others previously demonstrated antiproliferative effects of the PDE5 inhibitors tadalafil (7, 11) and vardenafil (11, 12) on prostatic stromal cells (Pr SC).In addition to these effects, we recently reported that tadalafil attenuated fibroblast-to-myofibroblast differentiation occurs via a two-step process that is initiated by changes in mechanical tension of the extracellular matrix that are transmitted to the fibroblast cytoskeleton via Ras homolog family member (Rho) A/Rho kinase (ROCK) signaling (13).TGFβ1 is considered to be a key inducer of pathogenic stromal reorganization, and we and others have demonstrated that TGFβ1 induces fibroblast-to-myofibroblast -differentiation (5–7).

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