Figure 3.

Cultured, Pax6-deficient astrocytes are immature and express neural stem cell markers. A, Immunostaining of GAFP, nestin, and prominin-1 (CD133) in WT and Sey/Sey astrocytes. Most WT astrocytes strongly express GFAP, whereas only a small number express nestin or prominin-1. In contrast, a smaller number of Sey/Sey astrocytes express GFAP, and a larger number strongly express nestin and prominin-1 compared with the WT astrocytes. Scale bar, 100 μm. The bottom two panels show the high-power images of prominin-1 expression in WT and Sey/Sey astrocytes. Scale bar, 25 μm. B, Cell lysates of cultured WT and Sey/Sey astrocytes were subjected to Western blot analysis with antibodies against GFAP, nestin, prominin-1, and Hsp90. The expression level of GFAP protein is slightly lower in Sey/Sey astrocytes than in WT astrocytes. In contrast, expression levels of nestin and prominin-1, markers for neural stem cells, are higher in Sey/Sey astrocytes than in WT astrocytes. C, Relative expression levels of GFAP, nestin, and prominin-1 protein normalized by Hsp90. The expression level of GFAP protein in Sey/Sey astrocytes is 0.73-fold lower than that in Sey/Sey astrocytes. The expression level of nestin and prominin-1 protein in Sey/Sey astrocytes is 1.61- and 6.2-fold higher than that in Sey/Sey astrocytes, respectively. The results are presented as the mean ± SD of samples (n = 3; p < 0.05).