Fig 1: (A) Extracellular matrix (ECM) contraction assay. Human colonic CCD-18Co fibroblasts were embedded in 100 µL/well of Matrigel (Corning #356234) and covered by 1 mL/well serum-free minimal essential medium (MEM). TGF-ß1 (10 ng/mL) and elafin (1 µg/mL) were added to the medium on day 0. The diameter of the Matrigel was measured on day 9. TGF-ß1 treatment significantly reduced the Matrigel diameter, indicating increased ECM stiffness. Shrinkage was unaffected by elafin. Results were pooled from 3 independent experiments. Ordinary one-way ANOVA with Tukey test. (B) Human colonic CCD-18Co fibroblasts were treated with elafin for 48 hours, followed by addition of MTS assay reagent (G5421; Promega, Madison, WI). Absorbance was determined at 490 nm. Results were pooled from 4 independent experiments. Ordinary one-way ANOVA with Tukey test. (C) Serum-starved primary human colonic epithelial cells were pretreated with TGF-ß1 (10 ng/mL) or CDSE (100 µg/mL) for 2 hours and then incubated with elafin for 48 hours. N-cadherin, ZEB1, and COL1A2 mRNA expression was determined by real-time reverse transcription polymerase chain reaction. TGF-ß1, but not CDSE, induced N-cadherin, ZEB1, and COL1A2 mRNA expression, unaffected by elafin. Results were pooled from 4 independent experiments. Ordinary one-way ANOVA with Tukey test.