ChABC exacerbated the epileptic seizure induced by PTZ.

<p>ChABC exacerbated the epileptic seizure induced by PTZ.</p

NBQX increased PNNs (<i>WFA</i>), tenascin-R, aggrecan and Neurocan in the medial prefrontal cortex in PTZ-treated rats.

<p>(A) Schemes of experimental schedules. (B) Numbers of <i>WFA</i>+ PNNs in mPFC of control and PTZ treatment, scale bar 50 μm, n = 6 per group. Representative <i>WFA</i>+ images of immunofluorescence staining are shown on the right. The data are expressed as mean ± SEM. The levels of (C) tenascin-R, (D) aggrecan and (E) Neurocan in mPFC are shown. Representative Western blot images are shown on the right. The data are expressed as a percentage of the values obtained for the rats treated with saline. *<i>p <</i> 0.01, different from corresponding saline groups, # <i>p</i> < 0.01, compared with PTZ group (<i>n =</i> 6).</p

Chronic PTZ treatment reduced PNNs (<i>WFA</i>), tenascin-R, aggrecan and Neurocan in the medial prefrontal cortex.

<p>(A) Schemes of experimental schedules. (B) Numbers of <i>WFA</i>+ PNNs in mPFC of control and PTZ treatment, scale bar is 50 μm, n = 6 per group. Representative <i>WFA</i>+ images of immunofluorescence staining are shown on the right. The data are expressed as mean ± SEM. (C) The levels of tenascin-R, aggrecan and Neurocan in mPFC are shown. Representative Western blot images are shown on the right. The data are expressed as a percentage of the values obtained for the rats treated with saline. *<i>p <</i> 0.01, different from corresponding saline groups (<i>n =</i> 6).</p

ChABC reversed the anti-epileptic effects of NBQX in PTZ-induced seizures.

<p>ChABC reversed the anti-epileptic effects of NBQX in PTZ-induced seizures.</p

NBQX decreased seizures induced by PTZ on latency to seizure (s), duration of the minor seizure onset (s), duration of the major seizure onset (s), and scores for the severity of seizures in rats.

<p>NBQX decreased seizures induced by PTZ on latency to seizure (s), duration of the minor seizure onset (s), duration of the major seizure onset (s), and scores for the severity of seizures in rats.</p

PTZ induced significant changes on latency to seizures (s), duration of the minor seizure onset (s), duration of the major seizure onset (s), and scores for the severity of seizures in rats.

<p>PTZ induced significant changes on latency to seizures (s), duration of the minor seizure onset (s), duration of the major seizure onset (s), and scores for the severity of seizures in rats.</p

Expression of FoxQ1 and NRXN3 in human normal brain and glioma tissues.

<p><b>A</b>, <i>NRXN3</i> and <i>FoxQ1</i> mRNA expression by RT-qPCR. The mRNA expression was analyzed in 30 matched primary glioblastoma tissues and the adjacent normal brain tissues. <b>B</b>, FoxQ1 expression levels correlated negatively with NRXN3 expression levels in glioblastoma samples (Pearson's correlation test <i>r</i> = −0.373; <i>P</i> = 0.042). <b>C,</b> Expression of FoxQ1 and NRXN3 protein in primary glioblastoma tissues and the adjacent normal brain tissues. Normal (N) and tumor (T) samples were analyzed by western blot. β-actin used as the loading control.</p

The NRXN3 as a transcriptional target of FoxQ1.

<p><b>A</b>, Sequence and position of putative FoxQ1 binding sites on the NRXN3 promoter. <b>B</b>, ChIP assays were done with U-87MG cells. Chromatin fragments of the cells were immunoprecipitated with anti-FoxQ1 antibody or negative control IgG (middle) and subjected to PCR. We subjected 1% of the total cell lysates to PCR before immunoprecipitation as inputs. <b>C</b>, schematic structure of the NRXN3 promoter. The sequence of the FoxQ1 binding sites are shown in both wild-type (WT) and mutant (Mut) forms. <b>D+E</b>, Luciferase activity with or without mutations in NRXN3 promoter. U-87MG cells were transfected with the wild-type NRXN3 promoter or its mutants (D). SW1088 cells were co-transfected with the wild-type NRXN3 promoter or its mutants and pcDNA3.1-FoxQ1 (E). Luciferase activities were then determined. Three independent experiments were conducted. * <i>P</i><0.05</p

FoxQ1 suppress the NRXN3 expression in human glioma cell lines.

<p><b>A</b>, Determination of FoxQ1 and NRXN3 expression in human glioma cell lines and normal human astrocytes using RT-qPCR (lower) and Western blot (upper). <b>B</b>, Up-regulation of NRXN3 mRNA and protein expression by overexpressing FoxQ1. FoxQ1 and NRXN3 expression levels in parental, control, SW1088-FoxQ1 cells by RT-qPCR (lower) and Western blot (upper). <b>C</b>, Down-regulation of NRXN3 mRNA and protein expression by depletion of FoxQ1 expression. FoxQ1 and NRXN3 expression in parental, control, and U-87MG-RNAi cells by RT-qPCR (lower) and Western blot (upper). <b>E+F</b>, Effect of FoxQ1 on NRXN3 promoter activity. Repression of the NRXN3 promoter in SW1088-FoxQ1 cells (E) and transactivation of the NRXN3 promoter in U-87MG-RNAi cells (F). Inhibition was calculated as a percentage relative to U-87MG cells and activation was calculated relative to SW1088 cells. Three independent experiments were conducted.</p

Down-regulation of NRXN3 rescues the malignant phenotype of FoxQ1 down-regulated glioma cells <i>in vitro</i> and <i>in vivo</i>.

<p>A, Western blot (upper) and RT-qPCR (lower) analyses of FoxQ1 and NRXN3 expression in stable NRXN3-rescued U-87MG cells. B, Cells as in (A) were cultured in 96-well plates and analyzed by MTT assay. Cell proliferation curves were shown in 9 days. Three independent experiments were conducted. C, Cells as in (A) were examined for cell migration motility in 24-well plates with transwell chambers. Migrated cells were stained with crystal violet and counted under a light microscope. Three independent experiments were conducted. D, Glioma cells (1×10⁶) were implanted intracranially into nude mice. Mice were euthanized when they were moribund or on day 90. * Incidence: number of mice with tumor/number of mice injected. E, Kaplan-Meier estimates of overall survival time in nude injected with glioma cells. *<i>P</i><0.05.</p