Figure 5

TGF-β1-pulsed ECM influenced the myofibroblast phenotype. Untreated fibroblasts were seeded onto decellularised TGF-β1-pulsed ECM. Classic myofibroblast markers were assessed 7 days post-culture. (a) Normalised densitometric SDS-PAGE analysis of the 24 h collagen secretion rate; (b) densitometric analysis of α-SMA normalised to β-actin expression; representative (c-f) corresponding SDS-PAGE gels and immunoblots of M1 and M7 ECM; (g) immunofluorescence images showing presence and distribution of α-SMA (red); F-actin (phalloidin, green) and nuclei stained with DAPI (blue). TCP control, matrix control and fibroblasts reseeded onto 4 h and 2 × 4 h TGF-β1-pulsed ECM images are modified to highlight α-SMA expression. Scale bars = 200 μM. *P <0.05 versus respective untreated controls. Data are represented as mean ± SD, calculated from three independent studies in triplicate, and expressed as fold changes over respective TCP controls. TGF-β1 ECM: ‘+’ denotes a 4 h and ‘++’ 2 × 4 h TGF-β1 pulse(s) on decellularised ECM. TGF-β1 cells: ‘+’ denotes a 24 h TGF-β1 pulse on reseeded fibroblasts. α-SMA, α-smooth muscle actin; ECM, extracellular matrix; SD, standard deviation; TCP, tissue culture plastic; TGF-β1 transforming growth factor-β1.