Figure 3. Analyses of astrocytes subjected to pressure. Representative fluorescence microscopic images of astrocytes are shown. About 3,000 isolated rat brain cortex astrocytes were subjected to pressures. Astrocytes were stained with rhodamine-phalloidin to determine the integrity of the actin cytoskeleton after 3 h of pressure treatment at the indicated pressure and a following 16 h recovery period: control cells (A) without pressure treatment, cells subjected to 25 mmHg (B), cells subjected to 100 mmHg (C), and cells subjected to 150 mmHg (D). All these cells were subjected to fluorescence microscopy with iso[4]LGE₂–protein adduct antibody after pressure treatment and control cells that did not go through any pressure treatment. E: Western blot analysis for iso[4]LGE₂ modification is displayed. Astrocytes were subjected to pressures (25–150 mmHg) for a period of 3 h at 37 °C. Bottom panel was probed with anti-GAPDH antibody.