Network of the genes turned on in PAH.

<p>(A) Top-scoring network derived from the 417 genes dramatically increased in PAH, despite their extremely low expression (<2 RPKM) in either normal adult heart or exercise-trained heart. The red-colored nodes represent up-regulated genes. (B) The histograms for the reads that were mapped by the UCSC isoforms of <i>Plk1</i>, <i>Foxm1</i> and <i>E2f1</i> in the different animal models. The blue lines underneath represent UCSC gene structures and the boxes show the exons of the genes. Expression of the 3 genes is only dominant in TAC. (C) The predicted motifs enriched in 1,000 bp upstream of 417 genes. The motifs shown on the bottom were predicted by MEME and the motifs shown above are the consensus motifs for the TFs (FOXM1 and PU.1). Y-axis indicates the amount of information at each position in the motif. The heatmaps (unit: RPKM) show the expression levels of the predicted TFs in sham (Sh), TAC (T), Se (Sedentary) and E (exercise). (D) Degree of expression (unit: RPKM) of the known targets of FOXM1 in the animal models are shown along with the previous evidence (PMID).</p

Experimentally verified isoforms alternatively spliced in PAH and PHH.

<p>(A) The histograms of mapped reads for 8 genes. The red asterisks shown at UCSC isoforms (blue lines underneath) indicate the exons (shown as blue boxes) alternatively spliced in hypertrophic signal-specific manners. The degree of expression is shown as vertical length. (B) Distribution of UCSC isoforms for the matching 8 genes characterized in different hypertrophy models. Different distribution of isoforms in either PAH or PHH was analyzed using NEUMA. The average RPKM values are shown with standard errors (N = 3). (C) Experimental confirmation of exon variants for the matching 8 genes using RT-PCR. For each RT-PCR results, the amplified regions with the specific primers are described. The detailed information for the primers used for the detection of the exons is available in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035552#pone.0035552.s007" target="_blank">Table S2</a>. <i>Fhl1</i>, <i>Rcan1</i>, <i>Ndrg2</i>, <i>Synpo</i> and <i>Ttll1</i> were experimentally confirmed in PAH and the splicing pattern of <i>Cxxc5</i> exon variants was confirmed in PHH. For <i>Egfl7</i> and <i>Tmpo</i>, the opposite splicing patterns were confirmed for both PAH and PHH. The isoform distributions of <i>Ttll1</i> and <i>Cxxc5</i> were not analyzed, since the UCSC isoforms for <i>Ttll1</i> is not reported and there were no mappable reads for <i>Cxxc5-2</i>. The exon numbers amplified are indicated on the right. For instance, E5∼6 means exon 5 and 6 were amplified with specific primers shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035552#pone.0035552.s007" target="_blank">Table S2</a>. For <i>Ndrg2</i>, two isoforms (E2∼5, +E3 and E2∼5, −E3) were shown in the same gel.</p

Critical pathways associated with DEGs and exon variants changed in PAH or PHH.

<p>Significantly enriched pathways (<i>p</i><0.05) with at least 5 DEGs or 5 exon variants for PAH and PHH are shown. Color intensity represents degree of enrichment (−log₁₀[<i>p</i>-value]). The significant pathways were categorized. (Group I) pathways for muscle contraction and metabolism. (Group II) pathways mainly for immune and cell cycle. (Group III) pathways for autoimmunity. (Group IV) pathways for cell signaling. (Group V) pathways mainly for cardiac diseases.</p

Platinum-Free Cathode for Dye-Sensitized Solar Cells Using Poly(3,4-ethylenedioxythiophene) (PEDOT) Formed via Oxidative Molecular Layer Deposition

Thin ∼20 nm conformal poly­(3,4-ehylenedioxythiophene) (PEDOT) films are incorporated in highly conductive mesoporous indium tin oxide (m-ITO) by oxidative molecular layer deposition (oMLD). These three-dimensional catalytic/conductive networks are successfully employed as Pt-free cathodes for dye-sensitized solar cells (DSSCs) with open circuit voltage equivalent to Pt cathode devices. Thin and conformal PEDOT films on m-ITO by oMLD create high surface area and efficient electron transport paths to promote productive reduction reaction on the PEDOT film. Because of these two synergetic effects, PEDOT-coated m-ITO by oMLD shows power conversion efficiency, 7.18%, comparable to 7.26% of Pt, and higher than that of planar PEDOT coatings, which is 4.85%. Thus, PEDOT-coated m-ITO is an exceptional opportunity to compete with Pt catalysts for low-cost energy conversion devices

Cells at different developmental stages were isolated, boiled in 3% SDS with 5% β-mercaptoethanol, subjected to SDS-PAGE and blotted with the specific antibodies

TC, testicular (spermatogenic) cells; TS, testicular sperm; S, mature sperm from the epididymis and vas deferens.<p><b>Copyright information:</b></p><p>Taken from "Characterization of eight novel proteins with male germ cell-specific expression in mouse"</p><p>http://www.rbej.com/content/6/1/32</p><p>Reproductive biology and endocrinology : RB&E 2008;6():32-32.</p><p>Published online 24 Jul 2008</p><p>PMCID:PMC2500023.</p><p></p

(A) Acrosome reaction of sperm from the epididymis and vas deferens was induced by calcium ionophore A23187

Acrosome-intact and -reacted sperm were subjected to western blot anaylsis with the anti-Shsp1/Mm.87328 antibody. AI, acrosome-intact sperm; AR, acrosome-reacted sperm. (B) Sperm from the epididymis and vas deferens were treated with 1% Triton X-100 and urea with different concentrations (2, 3, 4 and 6 M). Soluble and insoluble fractions after centrifugation of the treated sperm were subjected to western blot anaylsis with the anti-Sfap1/Mm.386907 and anti-Sfap2/Mm.157049 antibodies. Sup, supernatant after centrifugation; Pt, pellet after centrifugation.<p><b>Copyright information:</b></p><p>Taken from "Characterization of eight novel proteins with male germ cell-specific expression in mouse"</p><p>http://www.rbej.com/content/6/1/32</p><p>Reproductive biology and endocrinology : RB&E 2008;6():32-32.</p><p>Published online 24 Jul 2008</p><p>PMCID:PMC2500023.</p><p></p

miR-185 Plays an Anti-Hypertrophic Role in the Heart via Multiple Targets in the Calcium-Signaling Pathways

<div><p>MicroRNA (miRNA) is an endogenous non-coding RNA species that either inhibits RNA translation or promotes degradation of target mRNAs. miRNAs often regulate cellular signaling by targeting multiple genes within the pathways. In the present study, using Gene Set Analysis, a useful bioinformatics tool to identify miRNAs with multiple target genes in the same pathways, we identified <i>miR-185</i> as a key candidate regulator of cardiac hypertrophy. Using a mouse model, we found that <i>miR-185</i> was significantly down-regulated in myocardial cells during cardiac hypertrophy induced by transverse aortic constriction. To confirm that <i>miR-185</i> is an anti-hypertrophic miRNA, genetic manipulation studies such as overexpression and knock-down of <i>miR-185</i> in neonatal rat ventricular myocytes were conducted. The results showed that up-regulation of <i>miR-185</i> led to anti-hypertrophic effects, while down-regulation led to pro-hypertrophic effects, suggesting that <i>miR-185</i> has an anti-hypertrophic role in the heart. Our study further identified <i>Camk2d</i>, <i>Ncx1</i>, and <i>Nfatc3</i> as direct targets of <i>miR-185</i>. The activity of Nuclear Factor of Activated T-cell (NFAT) and calcium/calmodulin-dependent protein kinase II delta (CaMKIIδ) was negatively regulated by <i>miR-185</i> as assessed by NFAT-luciferase activity and western blotting. The expression of phospho-phospholamban (Thr-17), a marker of CaMKIIδ activity, was also significantly reduced by <i>miR-185</i>. In conclusion, <i>miR-185</i> effectively blocked cardiac hypertrophy signaling through multiple targets, rendering it a potential drug target for diseases such as heart failure.</p></div

<i>miR-185</i> negatively regulates NFAT activity in NRVMs.

<p>(A) Schematic representation of the luciferase reporter construct driven by nine tandem NFAT binding sites. (B) Relative luciferase activity in NRVMs transfected with NC or <i>miR-185</i> inhibitor. NRVMs were stimulated by ET-1 (10 nmol/L) for 24 h. pRL-TK was transfected for normalization and as an internal control for transfection efficiency. (C) 72 h after transfection of NC or <i>miR-185</i> mimic, the level of p-NFATC3 and total NFATC3 were analyzed by western blotting. α-tubulin was used as a loading control. Representative western blots (left) and quantified western blots (right). The data are expressed as mean ± SEM of more than three independent experiments; *<i>P</i> < 0.05, **<i>P</i> < 0.001.</p

<i>miR-185</i> negatively regulates the activity of CaMKIIδ in NRVMs.

<p>(A and B) 72 h after transfection of <i>miR-185</i> mimic or <i>miR-185</i> inhibitor, the level of p-CaMKIIδ and total CaMKIIδ were analyzed by western blotting. GAPDH was used as a loading control. (C) Western blotting showing p-PLB (Thr-17) and PLB protein expression in NRVMs transfected with NC or <i>miR-185</i>. 24 h after transfection, NRVMs were stimulated with ET-1 (10 nmol/L) for 48 h. The blots were stripped for 30 min and reprobed with PLB for loading control. The data are expressed as mean ± SEM; *<i>P</i> < 0.05, **<i>P</i> < 0.001, N = 3.</p

Testes from mutant mice (A) and a Sertoli cell line, TM4 (B), were subjected to protein extraction and western blot analysis using the specific antibodies

Total proteins from the testes were extracted by boiling the minced tissues in a protein sample buffer containing 3% SDS and 5% β-mercaptoethanol. ADAM2, with spermatogenic cell-specific expression, was included as a control. WT, wild type; TC, testicular (spermatogenic) cells.<p><b>Copyright information:</b></p><p>Taken from "Characterization of eight novel proteins with male germ cell-specific expression in mouse"</p><p>http://www.rbej.com/content/6/1/32</p><p>Reproductive biology and endocrinology : RB&E 2008;6():32-32.</p><p>Published online 24 Jul 2008</p><p>PMCID:PMC2500023.</p><p></p