Fig. 4. Effect of CARD-024 and substrate stiffness ECM protein and gene expression.
(A) Representative Western of αSMA protein expression in untreated or CARD-024 treated colonic myofibroblasts on low or high stiffness substrates. (B) Representative Western of pFAK protein expression in untreated or CARD-024 treated colonic myofibroblasts on low or high stiffness substrates. In (A) and (B) GAPDH protein expression is used as a control for protein loading. (C–H) QRT-PCR expression of fibrogenic (MLCK (C), ET-1 (D)or COX-2 pathway (PTGS2 (E), IL-1β (F)) genes in colonic myofibroblasts cultured on low or high stiffness substrates treated with CARD-024 (dark bars) compared to untreated cells (white bars). Asterisks denote comparisons between untreated/low stiffness and other groups. The horizontal reference line denotes baseline (untreated/low stiffness) expression. Pairwise comparisons between groups are denoted with brackets. * p <0.05, ** p <0.01, *** p <0.001.