Rho/MRTF-A-Induced Integrin Expression Regulates Angiogenesis in Differentiated Multipotent Mesenchymal Stem Cells
MRTF-A is essential for the VEGF-induced angiogenesis of differentiated MSCs and integrin α1, α5, and β1 expression. (a) After transfection with sh-MRTF-A or pSUPER (control), MSCs were cultured with EC differentiation medium for 3 d. MTT assay was performed to test the viability of MSCs. (b and c) MSCs were differentiated into ECs by treatment with VEGF for 7 d and siRNA-mediated knockdown experiments were then performed in the MSCs-derived ECs. The migratory ability of MSC-derived ECs transfected with si-MRTF-A or control siRNA was determined via wound healing (b) and transwell chamber assays (c). (d) Cell migration was quantified by calculating relative cell numbers. , . (e) After differentiation and siRNA or shRNA-mediated knockdown, the cells were cultured in matrigel for 7 d. Morphological changes in differentiated cells transfected with siMRTF-A or sh-MRTF-A were observed. (f) Capillary-like structures were quantified by measuring the polygonal network. , . (g) The expression of integrins α1, α5, and β1 was estimated by qPCR. ; , . (h) The expression of integrins α1, α5 and β1 was estimated by western blot. (i) The relative quantification of the protein expression. ; , .