The different microsatellite loci analyzed.

The different microsatellite loci analyzed.</p

Results for the hNSCs lines.

Results for the hNSCs lines.</p

Open field test and neuronal loss in ischemic brain areas at day 14.

<p> Mice receiving NC(4 h), but not those receiving NC(7 d) show a reversal of ischemia-induced impairment in number of rears and objects (A, n = 10) and a significant reduction in neuronal loss in striatum and cortex (B) compared to those receiving PBS (n = 6). Data are expressed as mean±SEM. One-way ANOVA: (A) rears F: 11.46, p<0.0001; (A) objects F: 5.38, p = 0.0056; (B) striatum F: 31.67, p<0.0001; (B) cortex F: 5.06, p = 0.0256. <i>Posthoc</i> Tukey test: °° p<0.01, °p<0.05 vs isch/PBS, ** p<0.01, * p<0.05 vs controls; §§§ p<0.001, § p<0.05 vs isch/NC(4 h).</p

mRNA expression of cytokines and trophic factors in ischemic cortex at day 1.

<p>Data are expressed as fold of induction compared to sham/PBS group (mean±SEM, n = 6). Two-way ANOVA: SDF-1α F: 97.52, p<0.0001; TGF-β1 F: 31.54, p = 0.0001; VEGF-A F: 24.14, p = 0.0004; IGF-1 F: 28.21, p = 0.0002; BDNF F: 19.34, p = 0.0009. <i>Posthoc</i> Bonferroni test: **p<0.01, ***p<0.001 vs sham/PBS; °°°p<0.001 vs isch/PBS; ### p<0.001 vs sham/NC(4 h).</p

Experimental design.

<p>NC were infused icv either 4 h (A) or 7 d (B) after ischemia. Histology, immunohistochemistry, confocal microscopy, real time PCR, behavioural test and neuronal count experiments were performed at the time points indicated.</p

Open field test and neuronal loss in ischemic brain areas at day 7.

<p>Mice receiving NC(4 h) show a reversal of ischemia-induced impairment in number of rears and objects, (A, n = 10) and a significant reduction in neuronal loss in striatum and cortex (B) compared to those receiving PBS (n = 6). Fibroblast infusion 4 h after ischemia does not significantly affect open field impairment and neuronal loss (n = 7). Data are expressed as mean±SEM. One-way ANOVA: (A) rears F: 6.122, p = 0.0002; (A) objects F: 16.10, p<0.0001; (B) striatum F: 17.67, p = 0.0002; (B) cortex F: 37.01, p<0.0001. <i>Posthoc</i> Tukey test: °° p<0.01, °p<0.05 vs isch/PBS; ** p<0.01 *p<0.05 vs controls; §§§ p<0.001 vs isch/NC(4 h).</p

Distribution of NC(4 h) in ischemic and sham-operated mice at days 1, 7 and 14.

<p>NC were infused icv in the lesioned side. Drawings represent typical results and are based on n = 5 brains for each experimental condition.</p

Confocal analysis of immunoreactivity of NC(4 h) - green - and nestin, GFAP or NG-2 - red - at day 1.

<p>Colocalization (arrows) can be observed between NC(4 h) and nestin, NC(4 h) and GFAP, NC(4 h) and NG-2. Images are taken in striatum, in proximity of the ventricular wall where most cells can be found at this time point. Bar: 25 µm.</p

Primers used for quantitative real-time polymerase chain reaction analysis of SDF-1α, BDNF, IGF-1, VEGF-A, TGF-β1, β-actin transcript levels.

a<p>forward</p>b<p>reverse</p

Quantification of microglia/macrophage activation.

<p>The area occupied by CD11b immunopositive cells was measured in striatum of sham-operated or ischemic mice receiving PBS, NC(4 h) or NC(7 d) at different time points (see Experimental design, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000373#pone-0000373-g001" target="_blank">Fig. 1A and B</a>). Data are expressed as mean±SEM (n = 4–5). Two-way ANOVA: (A) F: 58.05, p<0.0001; (B) F: 10.19, p = 0.0002. <i>Posthoc</i> Bonferroni test: *p<0.05, **p<0.01, ***p<0.001 vs sham/PBS; °°p<0.01, °°°p<0.001 vs isch/PBS, ##p<0.01, ###p<0.001 vs sham/NC.</p