An apoptotic cell generates high levels of S1P, but does not extrude in the presence of the S1P2 antagonist JTE-013. An HBE monolayer was treated with short-wave UV to induce apoptosis in the presence of JTE-013 and then stained with anti-S1P (green), Alexa Fluor 568–phalloidin for actin (red), and DRAQ5 (blue). Confocal micrographs were obtained using a microscope (TCS SP5; Leica) and Z series were reconstructed into 3D images using ImageJ and MetaMorph.
