All lanes : Anti-S1P antibody (ab59869) at 1/1000 dilutionLane 1 : Tissue lysate prepared from rat liver (1:4) at 20 µlLane 2 : Tissue lysate prepared from mouse liver(10mg/ml) at 20 µlLane 3 : Whole cell lysate prepared from human hepatoma SK-HEP-1 cells (1:4) (serum free) at 10 µlLane 4 : Whole cell lysate prepared from human hepatoma SK-HEP-1 cells (1:4) (treated with TPA) at 10 µlLane 5 : Whole cell lysate prepared from human hepatoma SK-HEP-1 cells (1:4) (treated with IL1-beta) at 10 µlLane 6 : Whole cell lysate prepared from human hepatoma SK-HEP-1 cells (1:4) (treated with TNF-alpha) at 10 µlLane 7 : Whole cell lysate prepared from human hepatoma SK-HEP-1 cells (1:4) (treated with RA) at 10 µl