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. 2009 Feb 28;41(2):116–125. doi: 10.3858/emm.2009.41.2.014

Figure 4.

Figure 4

Effects of VPA on neural differentiation and activation of Ras-ERK pathway in NSCs. (A) NSCs were either not treated or treated with 1 mM VPA for different time periods (0, 15 min, 2 h, 12 h or 48 h). The mRNA level of the BMP4, EGFR or HPRT were measured by RT-PCR, followed by agarose gel electrophoresis. (B) NSCs were either not treated or treated with 1 mM VPA for 48 h, and the levels of β-catenin, BMP4, pan-Ras, p-ERK, Tuj1, EGFR or β-actin were monitored by immune blot analyses. (C-D) NSCs were grown in the bFGF-containing medium (10 ng/ml). The cells were treated with 1 mM VPA for 48 h, together with or without 150 ng/ml of recombinant noggin. (C) Micrographs were taken using an ECLIPSE TE2000-U fluorescent microscope at ×400 magnification. (D) Pan-Ras, p-ERK, Tuj1, EGFR or β-actin were monitored by immune blot analyses. (E-F) NSCs were transfected with 2 µg of pcDNA3.0, dn-Tcf-pcDNA3.0, or dn-Ras in a 60 mm dish, and treated or non-treated with 1 mM VPA in the presence of bFGF (10 ng/ml) for 48 h. The levels of BMP4, pan-Ras, p-ERK, Tuj1 or β-actin were determined also by immunoblot analyses.