Figure 1. Effects of aging and L-CS on resting calcium (Ca2+) levels in cortical neurons.(A) The pictogram illustrates the experimental paradigm employed in the study. (B) Representative brightfield micrographs of control and aged neuronal cultures treated either with L-CS or vehicle (scale bar 100 µm). Please, note that aged cultures are devoid of signs of neuronal death. (C) Bar graphs depict the relative abundance of ceramides in vehicle- and L-CS-treated aged neurons (n=3 for both conditions). (D) Representative fluorescent micrograph of a fura-2-loaded cultured cortical neuron (the image reports dye emission when excited at 380 nm, scale bar 25 µm). (E) Bar graphs depict dendritic Ca2+levels of vehicle- or L-CS-treated control neurons (vehicle: n=102 proximal and n=85 distal dendrites from 43 neurons; L-CS: n=115 proximal and n=84 distal dendrites from 38 neurons; p>0.05). (F) Bar graphs depict dendritic Ca2+levels of vehicle- or L-CS-treated aged neurons (vehicle: n=182 proximal and n=156 distal dendrites from 40 neurons; L-CS: n=177 proximal and n=155 distal dendrites from 44 neurons; p>0.05). (G) Bar graphs depict somatic Ca2+levels of vehicle- or L-CS-treated control and aged neurons (ControlVeh: n=1357 cells and ControlL-CSn=1015; AgedVehn=539 cells and AgedL-CSn=497 cells obtained from 10-23 independent experiments). In C and E-F means were compared by unpaired Student t-test. In G means were compared by two-way ANOVA followed by Tukey post-hoc test. * indicates p<0.05, *** p<0.001.