Confocal image of Brdu (red) and Nestin (green) co-immunofluorescence staining in the ipsilateral SVZ 7 days after surgery.

<p>Panels A–C show staining of Brdu⁺/Nestin⁺ positive cells in the ipsilateral SVZ from Sham (A), MCAO (B), and rTMS (C) groups (Bar = 20 µm). (D) Quantification analysis of the number of Brdu⁺/Nestin⁺ positive cells in the ipsilateral SVZ 7 days after surgery. Brdu positive cells were labeled red (E), Nestin positive cells were labeled green (F) and Brdu⁺/Nestin⁺ positive cells were double-labeled (G), Bar = 20 µm. ^#<b>P<</b>0.001 for rTMS group versus Sham group; ^*<b>P</b> = 0.001 for rTMS group versus MCAO group. ^&<b>P</b> = 0.044 for MCAO group versus Sham group. SVZ, subventricular zone.</p

Infarct volume assessed by TTC staining 1 day after the tMCAO.

<p>(A) Position of SVZ in the coronal section of brain. Areas imaged for immunofluorescence studies are indicated by box. (B) Coronal brain section stained by TTC 1 day after tMCAO. The white areas without deep red-staining indicate ischemic areas. SVZ, subventricular zone.</p

Antagomir-25 injection resulted in decreased proliferation of NSCs 7days after tMCAO.

<p>Panels A–C show staining of Brdu⁺(red)/Nestin⁺(green) positive cells in the ipsilateral SVZ from rTMS (A), antagomir-25/rTMS (B), and control antagomir/rTMS (C) groups(Bar = 20 µm). (D) Quantification analysis of the number of Brdu⁺/Nestin⁺ positive cells in the ipsilateral SVZ at 7 days after tMCAO. ^#<b>P</b> = 0.006 versus rTMS group; ^*<b>P</b> = 0.013 versus Scr group. SVZ, subventricular zone.</p

Experimental protocol.

<p>Experimental protocol.</p

Effects of Ant-25 and Scr on p21 and p57 in the ischemic cortex 7 days after surgery.

<p>(A) Electrophoresis of p21, p57 and GAPDH on gel. (B) Relative levels of p21 and p57 protein from the different groups. ^#<b>P</b> = 0.018 compared to the rTMS group; ^*<b>P</b> = 0.012 compared to the Scr group.</p

Effects of Ant-25 and Scr on miR-106b∼25 in the ipsilateral cortex 7 days after surgery.

<p>(<b>A</b>–<b>C</b>) <b>A</b>) miR-93, <b>B</b>) miR-106b and <b>C</b>) miR-25 levels in the ischemic cortex after ICV injection of 1 nmol, 2.5 nmol and 4 nmol of Ant-25 or Scr. (D) qRT-PCR measurement of the miR-106b∼25 cluster in the ischemic cortex after ICV injection of 2.5 nmol of Ant-25 or Scr. ^#<b>P</b><0.001 for 2.5 nmol compared to the rTMS group; ^*<b>P</b><0.001 for 2.5 nmol compared to the Scr group. ^##P<0.001 for 4 nmol compared to the rTMS group.</p

Expression changes of miR-25 in the ipsilateral cortex 7 days after surgery.

<p>(A) Electrophoresis of miR-25 and U6 on gel. (B) Relative expressions of miR-25 in different groups. ^#<b>P</b><0.001 compared to the Sham group; ^*<b>P</b><0.001 compared to the MCAO group.</p

Neurobehavioral function was improved by rTMS after cerebral ischemia.

<p>NSSs were improved in MCAO rats treated with rTMS as compared with other groups. Data are presented as mean±SD. ^&P<0.001 versus Sham group. ^*P = 0.005 for MCAO group between day 7 and day 1. ^**P<0.001 for rTMS group between day 7 and day 1. ^#P = 0.019 between rTMS group and MCAO group 7 days after surgery.</p

Expression changes of p57 and PTEN in the ipsilateral cortex 7 days after surgery.

<p>(A) Electrophoresis of p57, PTEN and GAPDH on gel. (B) The ratio of the target genes to GAPDH in different groups. ^#<b>P</b> = 0.005 for p57 compared to the Sham group; ^#<b>P</b> = 0.004 for PTEN compared to the Sham group; ^*<b>P</b> = 0.007 for PTEN compared to MCAO group.</p

Genome-Wide Identification and Expression Profiling Analysis of <i>ZmPIN</i>, <i>ZmPILS</i>, <i>ZmLAX</i> and <i>ZmABCB</i> Auxin Transporter Gene Families in Maize (<i>Zea mays</i> L.) under Various Abiotic Stresses

<div><p>The auxin influx carriers auxin resistant 1/like aux 1 (AUX/LAX), efflux carriers pin-formed (PIN) (together with PIN-like proteins) and efflux/conditional P-glycoprotein (ABCB) are major protein families involved in auxin polar transport. However, how they function in responses to exogenous auxin and abiotic stresses in maize is largely unknown. In this work, the latest updated maize (<i>Zea mays</i> L.) reference genome sequence was used to characterize and analyze the <i>ZmLAX</i>, <i>ZmPIN</i>, <i>ZmPILS</i> and <i>ZmABCB</i> family genes from maize. The results showed that five <i>ZmLAXs</i>, fifteen <i>ZmPINs</i>, nine <i>ZmPILSs</i> and thirty-five <i>ZmABCBs</i> were mapped on all ten maize chromosomes. Highly diversified gene structures, nonconservative transmembrane helices and tissue-specific expression patterns suggested the possibility of function diversification for these genes. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the expression patterns of <i>ZmLAX</i>, <i>ZmPIN</i>, <i>ZmPILS</i> and <i>ZmABCB</i> genes under exogenous auxin and different environmental stresses. The expression levels of most <i>ZmPIN</i>, <i>ZmPILS</i>, <i>ZmLAX</i> and <i>ZmABCB</i> genes were induced in shoots and were reduced in roots by various abiotic stresses (drought, salt and cold stresses). The opposite expression response patterns indicated the dynamic auxin transport between shoots and roots under abiotic stresses. Analysis of the expression patterns of <i>ZmPIN</i>, <i>ZmPILS</i>, <i>ZmLAX</i> and <i>ZmABCB</i> genes under drought, salt and cold treatment may help us to understand the possible roles of maize auxin transporter genes in responses and tolerance to environmental stresses.</p></div