Figure 4
Adherent PD-neurospheres differentiate upon RA. A) Inverted light microscopic images of adherent PD-neurospheres (NT) or RA-treated for three days. Original magnification: 40X. The images shown are representative of three different experiments with similar results. B) Neurites length (μm) analysis. Distribution of the neurite outgrowth length as measured in adherent PD-neurospheres treated with RA for three days. Statistical significance: ***P ≤0.001. C) Representative fluorescent images of PD-neurospheres expanded for 12 passages and exposed to RA for one, three and seven days. Red = NF200 and βIII tubulin, Green = GFAP and NeuN immunostaining and blue = HOECHST. Scale bar: 10 microns. D) Distribution of intracellular DiOC5 fluorescence in adherent PD-neurospheres treated or not with RA for one, three and seven days expressed as % increase of treated cells versus untreated (black). Valinomycin treated cells (white) were used as hyperpolarised control. The result shown is representative of three different experiments overlapping. GFAP, glial fibrillary acidic protein; PD, placenta-derived; RA, retinoic acid.