Fig. 7. NE-induced translocation of cPLA₂ to the nuclear envelope depends on phosphorylation. VSMCs were treated with okadaic acid (OA; 10 µM), alkaline phosphatase (AP; 0.5 Unit ml^-1; loaded by reversible permeabilization with ß-escin), OA in combination with AP (OA+AP) or vehicle (VEH). (A) cPLA₂ translocation to the nuclear envelope in cells treated with 10 µM NE or its vehicle (V) was visualized by confocal microscopy (100x magnification). (B) Density of cPLA₂ fluorescence around the nuclear envelope (n=5). ^*Value significantly different from that obtained with V of NE (P<0.05). (C) cPLA₂ activity in VSMCs treated with OA, AP, OA+AP or VEH as described in Materials and Methods (n=5). (D) Phosphorylation of cPLA₂ as determined by ³²P incorporation in cells treated with OA, AP, OA+AP or VEH. A representative of three experiments performed with each agent or its vehicle on different batches of cells grown in 100 mm tissue culture dishes. (E) Density of cPLA₂ phosphorylation was quantified using NIH Image 1.62 (densitometric analysis of D).