Method: protocol for uniaxial cyclic stretch. (A) The elastomeric membrane of PDMS that serves as a substratum is first exposed to oxygen plasma and then coated with collagen type I. HUVECs are then seeded and cultured for 48 hours. Finally, uniaxial cyclic stretch is applied at a frequency of 1 Hz. IF followed by confocal acquisition. The automatic segmentation Cellpose was performed to quantify the parameters of interest of the individual cell (ROI). A typical confocal image of a HUVEC monolayer, in which DAPI (blue) and phalloidin (F-actin, red) are labeled, is shown. (B) Scheme depicting uniaxial cyclic stretch: triangular wave. Unstretched, 24 hours (blue); stretched at 10%, 6 hours (magenta); stretched at 10%, 24 hours (orange). Interrupted stretch: 24 hours, 10% stretch, followed by 6 hours discharge (yellow). Stretched 5%, 24 hours (green). All membranes have a flat substrate and cyclic stretch frequency set to f = 1 Hz. IF, immunofluorescence.
Figure 1.

Method: protocol for uniaxial cyclic stretch. (A) The elastomeric membrane of PDMS that serves as a substratum is first exposed to oxygen plasma and then coated with collagen type I. HUVECs are then seeded and cultured for 48 hours. Finally, uniaxial cyclic stretch is applied at a frequency of 1 Hz. IF followed by confocal acquisition. The automatic segmentation Cellpose was performed to quantify the parameters of interest of the individual cell (ROI). A typical confocal image of a HUVEC monolayer, in which DAPI (blue) and phalloidin (F-actin, red) are labeled, is shown. (B) Scheme depicting uniaxial cyclic stretch: triangular wave. Unstretched, 24 hours (blue); stretched at 10%, 6 hours (magenta); stretched at 10%, 24 hours (orange). Interrupted stretch: 24 hours, 10% stretch, followed by 6 hours discharge (yellow). Stretched 5%, 24 hours (green). All membranes have a flat substrate and cyclic stretch frequency set to f = 1 Hz. IF, immunofluorescence.

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