243 Fibroblast remodeling of extracellular matrix is directed by the fibrotic nature of the threedimensional microenvironment SUPPLEMENTARY FIGURES AND TABLES Supplementary Figure 1: Deposited collagen intensities in empty and fibroblast-seeded lung ECM-derived hydrogels. Control or IPF primary lung fibroblasts were seeded in control or IPF lung ECM-derived hydrogels and cultured for 7 or 14 days. Collagens were visualized using PicroSirius Red (PSR) staining on sections of paraffin-embedded fibroblast-seeded hydrogels and staining intensities were compared with their corresponding empty hydrogel samples. Day 7 responses with respect to A) weak B) moderate and C) strong PSR-stained intensities (arbitrary unit (au)). Day 14 responses with respect to D) weak E) moderate and F) strong PSR-stained areas (au). The dotted line shows the intrinsic difference between the empty IPF and control hydrogels. The estimate (± 95% confidence interval) shows the difference in the intensity of PSR-staining between the IPF and control hydrogels seeded with control (light blue, triangle) or IPF (dark blue, square) fibroblasts. P values below each fibroblast group represent the differences induced by fibroblasts in IPF versus control hydrogels compared to the intrinsic difference between empty IPF and control hydrogels. P-values above the estimates indicate the differences between the responses of IPF and control fibroblasts in the different hydrogels. Applied statistical test: mixed-model analysis. ns: not significant, IPF: Idiopathic pulmonary fibrosis. n=6 for fibroblast donors, 1 image per sample was analyzed. 9
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