Journal: BMC Neuroscience

Article Title: Poly(ADP-ribose) polymerase inhibitors activate the p53 signaling pathway in neural stem/progenitor cells

doi: 10.1186/s12868-016-0333-0

Figure Lengend Snippet: No induction of differentiation of NSPCs into neurons or glial cells by a PARP inhibitor. a The cells were triple-stained with Hoechst 33258 (nuclear staining), anti-nestin antibody (for NSPCs), and a cell-specific antibody: anti-Tuj-1 antibody (for neurons), anti-GFAP antibody (for astrocytes), or anti-CNPase antibody (for oligodendrocytes). Although the morphology of NSPCs was changed considerably after the addition of PJ34, the pattern of immunostaining was not changed. Differential interference contrast (DIC) images were on the left side. The positive controls for the antibodies used here are shown in the small images. b Quantitative mRNA analyses at 24 h after the addition of PJ34 ( shown in the upper row ) or 6 days after withdrawal of the growth factors EGF and FGF for differentiation into astrocytes ( shown in the lower row ) were performed for cell type-specific markers: Tuj-1 for neurons, nestin for NSPCs, GFAP for astrocytes, and CNPase for oligodendrocytes. Among these markers, GFAP was only upregulated by a few fold after the addition of 20 μM PJ34, while the expression of GFAP increased by more than 600-fold after in vitro differentiation of NSPCs into astrocytes. Data shown in ( b ) are expressed as the ratio of the mean value of the control ( vehicle alone ). Data represent the mean value ± SEM (n = 3).* p < 0.05 ( in the upper row ) by comparison against control using one-way ANOVA followed by Tukey’s post hoc test. † p < 0.01 and § p < 0.001 ( in the lower row ) by comparison against control using Student’s t test. Scale bars in ( a ), 100μm

Article Snippet: The cells were dissociated and suspended at a density of 2.0 × 10 6 cells in 100-mm dishes in 1× Dulbecco’s modified Eagle’s medium (DMEM)/F-12 neurosphere medium supplemented with B-27 (Gibco), 20 ng/mL human recombinant epidermal growth factor (EGF) (PeproTech), and 20 ng/mL human recombinant fibroblast growth factor (FGF)-basic (PeproTech).

Techniques: Staining, Immunostaining, Expressing, In Vitro