Mechanistic Studies
Primary SMCs are cultured in vitro under activating conditions to induce various trans-differentiations: oxLDL-loading to form foam-like cells, Ca/Pi treatment for osteogenic transition, PDGFB/TGFB1 for fibroblast-like phenotype, inflammatory cytokines to induce macrophage-like cells, cell stressors doxorubicine/irradiation to induce senescent subphenotype. State-of-the-art systems are then employed (e.g. live-cell imaging) that allow detailed cell phenotyping and siRNA is used for gene perturbations. Adhesion, migration, proliferation, apoptosis and contractility assays are performed to evaluate the mechanism of newly identified molecular targets in cell processes and basic molecular biology (Western blots, co-immunoprecipitations) for regulation of their signalling pathways.