S1P signaling regulates apoptotic cell efferocytosis and necroptotic cell extrusion. (A) Illustration of S1P signaling, and inhibitor targets. (B) Quantification of extruded versus retained pyroptotic and necroptotic cells in co-cultures treated with SKI-II or JTE-013. (C) Extrusion of pyroptotic and necroptotic cells over time in presence and absence of inhibitors. n = 3 independent experiments, number of cell death events is indicated above the bars. (D) S1P and LDH release from Caco-2 cells upon light-induced activation of optoCDEs. (E) Time-lapse images of opto-hCaspase-8-expressing Caco-2 cells undergoing apoptosis in presence of SKI-II or JTE-013. Scale bar, 20 µm. (F and G) Quantification of apoptotic cell retention versus extrusion in Caco-2 co-cultures treated with SKI-II and JTE013. (H) S1PR2 expression in Caco-2 cells transfected with non-targeting (NT) or anti-S1PR2 siRNA. (I) Extrusion of necroptotic and apoptotic cells over time co-cultured with bystander cells transfected with NT or S1PR2 siRNA. (J) Quantification of dying cell fates in MCF10A co-cultures. (K) Cell extrusion over time in MCF10A co-cultures treated with JTE-013 or SKI-II. Cells were photostimulated with blue light 8.7 mW/cm² every 180 s to activate optoCDEs. Mean ± SEM (C, G, I, and K) or mean ± SD (D and H). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (Mantel-Cox test). All data are representative of (E) or pooled from (B–D and F–K) three independent experiments, each done with triplicate technical replicates (n = 9). (D) Data points represent technical replicates.