FIGURE 2.

LCAD downregulation is associated with increase of NSC proliferation. The NSC line NS-TGFP was induced to differentiate for up to 72 h and total proteins were collected for immunoblotting analysis. NSCs were also transfected with either control or LCAD siRNAs, and collected for RT-PCR analysis to assess mRNA expression levels of proliferation, stemness and differentiation markers after 24 h. In addition, adult rats were treated with TUDCA or vehicle for 28 days using miniosmotic pumps, as described in Materials and Methods. Immunohistochemistry against LCAD (red) and NSC early differentiation marker DCX (green) was performed. (A) Representative immunoblots of total LCAD levels (top) and corresponding densitometry analysis (bottom) in NSCs throughout differentiation. Total LCAD levels were normalized to GAPDH. (B) mRNA expression levels of proliferation (ki67), stemness (Nestin) and differentiation (Map-2) markers in NSCs collected 24 h after siRNA transfection. Values were normalized to the internal standard HPRT. Data are expressed as mean ± SEM fold-change for at least three independent experiments. **p < 0.005 and *p < 0.05 from control (untreated cells). (C) Representative confocal images of LCAD- and DCX-positive cells in frontal sections of adult SVZ neurogenic niches in TUDCA-treated and untreated rats (n = 4). Nuclei were stained with Hoechst. Scale bars, 75 μm in SVZ image and 25 μm in representative image sections with or without TUDCA treatment.