Figure 1

Neurosphere cultures and immunocytofluorescence. For in vitro differentiation cells from the basal ganglia of 15.5 dpc C57BL/6 mice were cultured in neurospheres and dissociated after 7 days. FGF2 was withdrawn after 2.5 days and treatment started 1.5 days after plating. Cells were treated with TSA (10, 25 or 50nM) or BMP2 (10 ng/ml). RNA and proteins were isolated after 6, 12 and 24 h (A). For immunocytofluorescence (B,C), cultures were treated with vehicle (CTL), 50nM trichostatin A (TSA), 10 ng/ml recombinant BMP2 (BMP2), or both reagents (BMP2/TSA) for 24 hours before bFGF withdrawal. Cultures were fixed after 4.5 additional days and stained with the following antibodies: TuJ1 (B, green) to label newborn neurons, anti-GFAP (B, red) to label newborn astrocytes, or O4 (C, red, indicated with arrows) to label newborn oligodendrocytes. DAPI (blue) was used to stain nuclei. Scale bar = 50 (B) and 100 (C) μm.