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Here we unveil the molecular mechanism by which cell spreading and Rho A GTPase activity control FA formation through YAP to stabilize the anchorage of the actin cytoskeleton to the cell membrane.
This mechanism requires YAP co-transcriptional function and involves the activation of genes encoding for integrins and FA docking proteins.
To this end, we cultured adipose tissue-derived mesenchymal stem cells (AD-MSCs) onto fibronectin (FN)-coated elastically supported surfaces of different stiffness (28 and 1.5 k Pa) or onto glass surfaces coated either with FN or poly--lysine (PLL).
This experimental setting allowed us to finely tune FA number regardless of cell size.Cells were stained with anti-YAP (red), and anti-vinculin (green).Graphs: quantification of FA number, total FA area and YAP nuclear/cytoplasmic ratio in single AD-MSCs grown onto fibronectin-coated surfaces with increasing adhesion areas.Graphs: quantification of YAP nucleus/cytoplasm ratio and cell area in transfected cells.(d) Left: Annotation of micropattern properties (cell area, adhesion area), schematic side view of single cell onto fibronectin-coated micropatterned glass slides, schematic top view of fibronectin distribution in the micropatterns. Center: confocal images of single AD-MSCs grown onto micropatterns stained with anti-vinculin (green).
In fact, both YAP nucleus/cytoplasm ratio and the number of FAs increased steadily with the size of the cell.