Figure 3.

Temperature-induced stabilization of the cytoskeleton dramatically increases attachment strength. (A) The attachment strength of HT1080 cells to fibronectin in PBS+MgCa conditions is shown after treatment with DTSSP (60 min), PO (120 min), cytD (60 min), or lat A (30 min). Temperature during shear application was either 37°C or 8°C. (B) Representative fluorescence images showing DNA (blue), actin (red), and paxillin (green) of cells subjected to shear at 37°C (left) or 8°C (right) at indicated shear in PBS+MgCa conditions, relative to their condition attachment strength, where <τ₉₀ indicates that at least 90% of the cells remained attached, whereas >τ₅ indicates >95% have detached. (C) Quantification of detachment mechanisms using at least 100 cells (or footprints) per condition. Cells were characterized as Intact (e.g., B, top), partly peeled defined as nucleus present and paxillin puncta were observed at cells’ leading edge relative to the shear direction (e.g., B, middle left), Detached+FAs, only paxillin puncta resembling cellular footprints visible (e.g., B, bottom left), Detached+FAs+Actin, paxillin puncta resembling cellular footprints and actin visible. To see this figure in color, go online.