Figure 6. Western blot analyses for determining iso[4]LGE₂-modification of proteins in extracts from astrocytes treated with pyridoxamine. About 3,000 isolated rat brain cortex astrocytes were treated with freshly prepared 5–50 mM pyridoxamine (except for the control sample) in water as indicated. Astrocytes were subjected to treatment with pyridoxamine before or after pressure treatment (100 mmHg for 4 h followed by a recovery period of 16 h at 37 °C and at atmospheric pressure). Western blot analysis was performed with iso[4]LGE₂–protein adduct antibody after the transfer of 25 µg of total protein extracts onto a PVDF membrane. Bottom panel shows probing with anti-GAPDH antibody.