This record contains raw data related to the article "Influence of culture substrates on morphology and function of pulmonary alveolar cells in vitro" ABSTRACT: Cell’s microenvironment has been shown to exert influence on cell behavior. In particular, matrix stiffness and cell interaction strongly impacts cell morphology and function. The purpose of this study was to analyze the influence of different culture substrate materials on phenotype and functional properties of lung epithelial adenocarcinoma (A549) cells. A549 cells were seeded onto polyester (Thermanox™ Coverslips) and polydimethylsiloxane (PDMS, Elastosil® Film) substrates with elastic modulus of 1,350 ±290 and 0.85 ±0.01 MPa, respectively. The two substrates influenced cell morphology and the actin cytoskeleton organization. Further, the Yes-associated protein (YAP) and its transcriptional coactivator PDZ-binding motif (TAZ) were translocated to the nucleus in A549 cells cultured on the more rigid polyester substrate, yet it remained mostly cytosolic in cells on soft PDMS substrate. By SEM analysis, we observed that cells grown on Elastosil® Film maintained an alveolar Type II cell morphology. Immunofluorescence staining for surfactant-C revealing a high expression of surfactant-C in cells cultured on Elastosil® Film, but not in cells cultured on Thermanox™ Coverslips. A549 cells grown onto soft Elastosil® Film exhibited morphology and functionality that suggest retainment of alveolar epithelial Type II phenotype, while A549 cells grown onto more rigid substrates acquired an alveolar Type I phenotype.