Figure 6

Nolz1 acts as a neurogenic factor in primary LGE cultures. (A,B) Double fluorescent immunocytochemistry performed on primary E14.5 LGE cultures. Nolz1 is expressed in cells positive for the early neuronal marker Tuj1 (A). In contrast, Nolz1 is not detected in cells expressing the astroglial marker glial fibrillary acidic protein (GFAP) (B). White arrows show double positive cells, open arrows show single stained cells for Tuj1 or GFAP, and white arrowheads show single stained cells for Nolz1. Scale bar: 50 μm. (C,D) Over-expression of hNolz in LGE primary cultures increased the number of neurons at the expense of glial fates as shown by the increase in the number of Tuj1- or MAP2-positive cells and the complete blockade of astroglial cells. Note that all cells over-expressing hNolz are negative for the GFAP marker (C). (C) White arrows show single stained cells for Nolz1, white arrowheads show double positive cells and open arrows show single GFAP-positive cells. Scale bar: 50 μm. (D) The results represent the mean ± standard error of the mean from at least three independent samples at each condition. Statistical analysis was performed with the Student's t-test. *P < 0.05, ***P < 0.001 relative to EGFP control.