Figure 1
PtdCho and PtdEtn impact on neurosphere-derived cells differentiation. (a) Neurosphere-derived cells cultured during 3 days under differentiation condition (control) or in the presence of liposomes of PtdCho or PtdEtn were immunostained with antibodies against βIII-tubulin (green), glial fibrillary acid protein (GFAP) (red) or Olig-2 (green). Nuclei were counterstained with DAPI (blue). Pictures were taken with Nikon Model Eclipse 800 microscope and are representative of independent experiments conditions. (b–d) Graphs represent the percentage of neuronal (βIII-tubulin), astroglial (GFAP) and oligodendroglial (nuclear-Olig2) cells after 3 days under the indicated condition of differentiation. (e) Western blot analysis was performed for βIII-tubulin and γ-tubulin as a control. The gels/blots displayed here are cropped, and without high-contrast (overexposure). The full-length gels and blots are included in a Supplementary Information file. (f) Neurosphere-derived cells were treated with 50 μM of PtdCho for 1 hour and then the media was replaced for phospholipid-free media and incubated for 3 days. (g) Percentage of neuronal cells (βIII-tubulin positive cells) when PtdCho was added later on, after 24 h of culture and incubated for 3 days. Immunostained were performed after 3 days of culture in each assayed conditions. Graphs are representative of at least three independent experiments. Data were presented as mean ± SEM. ***p < 0.001; *p < 0.05.
