Effects of EA treatment on neurogenesis at 14 days after MCAO.

<p>Schematic diagram shows the regions of the hippocampus (★, in and around the hippocampus), SVZ (*, including the striatum) and cortex (#, the whole cortex) of the brain for immunohistochemical and further Western blot analysis (A). Photomicrograph (B) and its histogram for BrdU/Dcx double-positive cells in the hippocampus (C), SVZ (D) and cortex (E) of the brain. BrdU/Dcx double-positive cells were significantly increased by EA treatment in the hippocampus and SVZ of the ipsilateral hemisphere. ^#<i>P</i><0.05, ^##<i>P</i><0.01 and ^###<i>P</i><0.001 versus the contralateral hemisphere; *<i>P</i><0.05 and **<i>P</i><0.01 versus MCAO mice. Scale bars = 100 µm.</p

Schematic diagram for experimental procedures.

<p>All experiments were performed on the indicated day.</p

RT-PCR analysis for growth and neurotropic factors at 14 days after MCAO.

<p>Expression of each mRNA is expressed as a percentage of contralateral hemisphere of MCAO. Panel (A) its densitometric analysis (B). Panel represents a typical result from three independent experiments. EA treatment significantly increased the mRNA level of BDNF and VEGF in the ipsilateral hemisphere. *<i>P</i><0.05 and ***<i>P</i><0.001 versus MCAO mice.</p

Effects of EA treatment on neurogenesis in the whole brain at 26 days after MCAO.

<p>Serial sections of the whole brain showing the regions for immunohistochemical analysis (A, cresyl violet stain) and its analysis for double-positive cells BrdU with neuroblast marker Dcx (B) or neuronal marker NeuN (C). Photomicrograph represents a typical result in the cortex. The number of proliferated NSCs indicated by BrdU positive was increased in both hemispheres of MCAO mice and EA treatment significantly increased the number of these cells in the posterior region of the brain. Total number of BrdU and Dcx or NeuN double-positive cells was significantly increased by EA stimulation. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001 versus MCAO mice. Scale bars = 100 µm.</p

Western blot and immunohistochemical analysis for VEGF at 14 days after MCAO.

<p>Western blot (A) and its densitometric analysis (B) showed that EA treatment significantly improved the expression of VEGF in the ipsilateral hippocampus and cortex. Photomicrograph (C) and its histogram (D) showed that VEGF positive cells were significantly increased by EA treatment in the hippocampus and ipsilateral SVZ. ^#<i>P</i><0.05 and ^##<i>P</i><0.01 versus contralateral hemisphere; *<i>P</i><0.05 and ***<i>P</i><0.001 versus MCAO mice. Scale bars = 100 µm.</p

Western blot and immunohistochemical analysis for mBDNF at 14 days after MCAO.

<p>Western blot (A) and its densitometric analysis (B) showed that EA treatment significantly improved the expression of mBDNF in the ipsilateral hippocampus. Photomicrograph (C) and its histogram (D) showed that mBDNF positive cells were significantly increased by EA treatment in the ipsilateral hippocampus and SVZ. ^#<i>P</i><0.05 versus the contralateral hemisphere; *<i>P</i><0.05 and **<i>P</i><0.01 versus MCAO mice. Scale bars = 100 µm.</p

Immunohistochemical analysis for pPI3K in newborn cells at 14 days after MCAO.

<p>Number of pPI3K/BrdU double-positive cells was significantly increased by EA treatment in all regions examined. ^#<i>P</i><0.05 versus contralateral hemisphere; *<i>P</i><0.05 versus MCAO mice. Scale bars = 100 µm.</p

Time-course effects of 1-methoxyoctadecan-1-ol on calpain activation, STEP cleavage, and p38 MAPK phosphorylation after glutamate exposure.

<p>Cortical neurons were treated with glutamate (300 µM, 15 min) (A and C) and pretreatment with 1-methoxyoctadecan-1-ol (0.1 µg/ml, 24 h) followed by exposure to glutamate (B and D) and both were maintained in the original medium for the specified time. Equal amounts of proteins and each sample were subjected to Western blot assays using the indicated antibodies. Equal protein loading was confirmed by actin expression.</p

Effects of 1-methoxyoctadecan-1-ol on infarct volume, neurological evaluation, and wire-grip test in a photothrombotic ischemic model.

<p>Mice received intraperitoneal administration of DMSO or 1, 3/kg of 1-methoxyoctadecan-1-ol at 30 min before the ischemic insult. Representative photographs of coronal brain sections stained with TTC in vehicle (Veh)- and 1-methoxyoctadecan-1-ol-treated mice (A). White indicates the infarct area. Quantification of the infarct volume, neurological score, and wire-grip test (B). *<i>P</i><0.05, **<i>P</i><0.01 <i>vs</i>. vehicle group. Data are expressed as mean±SEM of three separate experiments.</p

Effects of 1-methoxyoctadecan-1-ol on calpain activation, STEP cleavage and p38 MAPK phosphorylation.

<p>The significant calpain1 activation (A), STEP cleavage (B) and p38 MAPK phosphorylation (C) were shown in the ipsilateral (Ipsil) cerebral hemisphere of photothrombotic ischemic mice compared with the contralateral (Con). **<i>P</i><0.01, ***<i>P</i><0.001, <i>vs</i>. vehicle group, Data are expressed as mean±SEM of three separate experiments.</p