Figure 2
PtdCho and PtdEtn modified the fate of post-mitotic cells. (a) Bright field images of neurosphere-derived cells obtained by time-lapse video-microscopy. The numbers on the right upper corner indicate the time of imaging (See also Supplementary Movie). (b) Post-imaging immunocytochemistry of tracked cells after a 3-day experiment using antibodies against the neuronal protein MAP2 (upper panel) or the glial protein GFAP (lower panel). Observe that cells shown in panel (a) and indicated by green and red arrows express MAP2, whereas the cell pointed by a blue arrow expresses GFAP. (c) Schematic representation of lineage trees reconstructed from time-lapse recording. Green arrow: progenitor cellss that generate two daughter cells that express βIII-tubulin. Red arrow: post-mitotic cells that express βIII-tubulin. Blue arrow: post-mitotic cells that express GFAP. (d) Percentage of cells undergoing cell division (progenitors) or not (post-mitotic cells) during the 3-days period of real-time observation. (e) Quantification of BrdU labeled cells in each indicated condition. The analysis is representative of three independent experiments. (f) Western blot analysis was used to investigate the amount of PCNA and γ-tubulin as a loading control, in total extract obtained from neurosphere-derived cells cultured under the indicated conditions. (g) Quantification of MAP2+ neurons generated from progenitor cells or (j) differentiated from post-mitotic cells present in the culture since the beginning of imaging. (m) Quantification of GFAP+ astrocytes differentiated from post-mitotic cells. (i) Quantification of cells generated from progenitor cells during the 3-days imaging period expressing neither MAP2 nor GFAP (“unlabeled”). (l) Quantification of unlabeled non-dividing cells. (h) Quantification of cell death among cells generated from progenitor cells during the 3-days of observation. (k) Quantification of cell death among non-dividing cells. Data were presented as mean ± SEM *p < 0.05; **p < 0.01.
