Y-27632 enhances the differentiation of P19 cells into ectoderm.

<p>P19 cell aggregates were treated with RA+LIF to induced neural-ectoderm differentiation. Total mRNA was collected, converted to cDNA and used in a PCR to examine GFAP expression, a marker of ectodermal derived neural cell types (A). GFAP levels increased with RA+LIF treatment in the presence or absence of Y-27632. Increased GFAP expression was also detected in the DMSO treated groups supplemented with Y-27632. Immunofluorescent detection of GFAP in undifferentiated (B) RA+LIF (C) RA+LIF and Y-27632 (D) or RA+LIF with Y-27632 pre-treated cells (E). qPCR with NF-68 primers showed an increase in expression in cells exposed to Y-27632 during differentiation (F). Immunofluorescent detection of NF-68 in undifferentiated cells or those exposed to the following: (G) RA+LIF (H) RA+LIF and Y-27632 (I) or RA+LIF with Y-27632 pre-treatment (J). FACS analysis demonstrates increased numbers of GFAP- (trend) and NF-68-positive cells with Y-27632 treatment during RA+LIF induced differentiation (K). Scale bar equals 50 µm, green staining represents the primary antibody, and blue staining is the nuclear dye TOTO-3. * Significance accepted at p<0.05.