Effect of apelin deficiency on disease progression of ALS mice.

<p>(A) Motor signs (hind limb tremors) were measured as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023968#pone-0023968-g003" target="_blank">Fig. 3B</a>. (B) Rotarod performance of SOD1^G93A littermates (open circles) and KO-SOD1^G93A (closed circles) mice was measured as described in the methods for 180 s. (C) The number of motorneurons was decreased in the lumbar spinal cord of KO-SOD1 ^G93A mice at 14 weeks-old (n = 3). Data represent mean ± SEM. *<i>p</i><0.05 vs. SOD1 ^G93A mice.</p

Effect of apelin and VEGF on oxidative stress-induced neurotoxicity in rat primary neurons.

<p>(A) Representative picture shows the expression of APJ and apelin mRNA in primary cortical and hippocampal neuronal cultures (representatively, C and H). Culture medium was replaced with neurobasal media without B27 supplement for 24 hours. (B) Cell viability of hippocampal neuronal cultures treated with indicated concentration of apelin for 24 hours. The cultures were treated with the indicated concentration of apelin (C) or with VEGF (50 ng/ml) and apelin (indicated concentration) (D) for 15 min prior to exposure to H₂O₂ (10 µM) for 24 hours. Cell viability was assessed by MTT assay (n = 4). *<i>p</i><0.05 vs. control. Data represent mean ± SEM.</p

APJ expression in the lumbar spinal cord of WT mice.

<p>Representative pictures of ventral horn show double immunostaining for APJ and NeuN (A–C) or GFAP (D–F) in the lumbar spinal cord. Scale bar = 200 µm (upper panels), 100 µm (lower panels).</p

Microglial activation in apelin deficient mice with mutant SOD1.

<p>Representative pictures show that Iba1-positive microglia in the lumbar spinal cord of SOD1 ^G93A (A) and KO-SOD1^G93A (B) mice. (C) The Iba1-positive area was increased in the lumbar spinal cord of KO-SOD1^G93A mice at 14 weeks-old (n = 3). Scale bar = 50 µm. *<i>p</i><0.05 vs. SOD1^G93A. Data represent mean ± SEM.</p

Time course of disease progression in SOD1^G93A mice monitored with three different tests.

<p>(A) Motor signs (hind limb tremors) were measured with the clinical scoring system (see methods). (B) Body weight was monitored twice a week. (C) The number of ChAT-positive cell bodies was decreased at end-stage disease for SOD1^G93A mice. **p<0.01 vs. wild-type mice. Data represent mean ± SEM.</p

Gene expression in lumbar spinal cord of SOD1^G93A mice.

<p>Temporal expression patterns of apelin (A) and APJ (B) in the lumbar spinal cord of wild-type (open column) and SOD1^G93A (closed column) mice were examined by real-time RT-PCR (n = 3–6). Data are mean ± SEM. **<i>p</i><0.01 vs. wild-type mice.</p