Research Paper Volume 11, Issue 16 pp 6336—6357

Inhibition of de novo ceramide biosynthesis affects aging phenotype in an in vitro model of neuronal senescence

Figure 2. Effects of aging and L-CS on intracellular Ca2+stores and NCX activity in cortical neurons.(A) Time course of CCCP-stimulated Ca2+release from mitochondria. Traces represent the average response to a 3 min exposure to 5 µM CCCP (ControlVeh: n=226 cells and ControlL-CSn=140; AgedVehn=91 cells and AgedL-CSn=67 cells obtained from 7-19 independent experiments). (B) Dot plots depict Ca2+peak obtained in the four study groups. (C) Dot plots depict Ca2+changes expressed as AUC (a.u.). (D) Time course of CPA-stimulated Ca2+release from the ER. Traces represent the average response to a 2 min exposure to 10 µM CPA (ControlVeh: n=50 cells and ControlL-CSn=33; AgedVehn=54 cells and AgedL-CSn=48 cells obtained from 3-4 independent experiments). (E) Dot plots depict Ca2+peak obtained in the four study groups. (F) Dot plots depict Ca2+changes expressed as AUC (a.u.). (G) Time course of NCX activity imaged by stimulating exchanger reverse operational mode (ControlVeh: n=163 cells and ControlL-CSn=122; AgedVehn=106 cells and AgedL-CSn=98 cells obtained from 2 independent experiments). (H) Dot plots depict Ca2+peak obtained in the four study groups. (I) Dot plots depict Ca2+changes expressed as AUC (a.u.). Means were compared by two-way ANOVA followed by Tukey post-hoc test. * indicates p<0.05, *** p<0.001.