Expression of Gli2 induces neurogenesis in P19 EC cells.

<p>(A–C): P19[Gli2] and P19[Control] cells were stained with Tuj1 and NF68 antibodies on day 6 or GFAP antibodies on day 10 of –RA differentiation. Nuclei were stained with Hoechst, scale bar is 30 µM. (<b>D</b>): Tuj1-, NF68- and GFAP-positive cells from (A)–(C) were counted in 10 random fields and expressed as a percentage of the total number of nuclei (10,000 cells; n = 4) (**p<0.01, n.s. = not significant). (<b>E</b>): NF68 immunoblot using total protein from day 6 differentiated P19, P19[Control] and two clonal populations of P19[Gli2] cell lines. P19 cells were differentiated in the presence of RA and served as a positive control. α-tubulin served as a loading control. (<b>F</b>): Total protein from –RA differentiating P19[Control] and P19[Gli2] cells was harvested on the days indicated, separated and immunoblotted with Gli2-specific antibodies. β-actin served as a loading control. Asterisk denotes non-specific binding of Gli2 antibodies.