21 research outputs found

    Early increase and later decrease in GSK-3β activity.

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    <p>Western blot of total GSK-3β and GSK-3β phosphorylation at ser9 (a: blot, b: quantification of phospho corrected for total GSK-3β). P-GSK-3β at Ser 9 inhibits phosphorylation of its substrate GS1 as depicted in Figure 5c. These data were derived from the kinomics chip. Data are presented as mean ± SD of n = 3.</p

    β-catenin levels decrease over time.

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    <p>Western blot (a) and quantification (b) of β-catenin. β-catenin protein levels decrease during <i>S. pneumoniae</i> pneumonia. Indirectly, represent a decrease of potentially active β-catenin. Data are represented as mean ± SD of n = 3.</p

    Clustering.

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    <p>Hierarchal clustering according to Johnsons of phosphorylation states of whole lung lysates <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0018519#pone.0018519-Johnson1" target="_blank">[18]</a> (a). Venn diagram of spots phosphorylated at measured time points. 3 hours is depicted in blue, 6 hours in yellow, 24 hours in green and 48 hours in red (b). Kinase activities spanning multiple time points (intermediate colors) are listed.</p

    Decrease in CDK activity with increasing pneumonia.

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    <p>Western blot (a) and quantification of all CDK phosphorylated substrates (b) revealed a decrease in CDK activity. Quantification was performed for all positive signals on the blot and corrected for β-actin. Data are represented as mean ± SD of n = 3.</p

    Provisional signal transduction scheme of active signaling pathways during pneumonia.

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    <p>Activation is depicted in green and inhibition in red, direction of events is calculated in relation to the noninfected control. Events from all timepoints (3, 6, 24 and 48 hours) were used to construct this scheme representing the entire host response dynamics of pneumococcal pneumonia. Important are the overall increment of chemotoxic stress and the initiation of the Th1 response. Spot numbers and activities (up ↑ or down ↓) are presented with corresponding kinases and timepoints. WNT signaling and the cell cycle are reduced throughout <i>S. pneumoniae</i> pneumonia.</p

    AMPK-α activity declines during pneumococcal pneumonia.

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    <p>Western blot analysis of phosphorylated AMPK-α (pAMPKα and unphosphorylated AMPK-α (b,c). During course of infection the ratio of pAMPK/AMPK decreased. A similar activity pattern was found in the kinomics chip (a). The bar graph shows quantification of the relative amounts of phospho-AMPK-α corrected for total AMPK-α. Data are presented as mean ± SD of n = 3.</p

    Cullin box domain promotes induction of <i>hes1</i> gene <i>in vitro</i>.

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    <p>nTera-d1 cells were co-transfected with <i>hes1-luciferase</i> (<i>hes1</i>) or <i>hes1-luciferase</i> lacking the conserved CSL-binding site (<i>hes1-RBPdel</i>) and <i>myc-tag</i> (MT) as a control, or myc-tagged <i>d-asb11</i> full length (MT-Asb11) or myc-tagged <i>asb11<sup>cul</sup></i> (MT-Asb11<sup>cul</sup>) cDNA. Hes1-dependent Notch activity was analyzed by luciferase measurement.</p

    <i>asb11<sup>cul</sup></i> presented altered expression of Delta-Notch pathway components.

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    <p>Wild type (<i>left panel</i>) and mutant (<i>middle panel</i>) embryos at 12 hpf were analyzed for WISH using probes against <i>her1</i>, <b>A</b>; <i>her4</i>, <b>B</b>; <i>her5</i>, <b>C</b>; <i>notch3</i>, <b>D</b>; <i>deltaD</i>, <b>E</b>; <i>and deltaA</i>, <b>F</b>. (<b>G</b>), Higher magnification shows detailed analysis of <i>deltaA</i> expression. (<i>left</i>) Graphs quantify the mRNA expression levels.</p

    Cullin box is essential for DeltaA degradation and for maintaining a cell proliferating state <i>in vivo</i>.

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    <p>(<b>A</b>) Zebrafish embryos were injected with Myc-tagged <i>deltaA</i> (MT-DeltaA) and <i>d-asb11</i> (Asb11) or <i>asb11<sup>cul</sup></i> (Asb11<sup>cul</sup>) mRNA at one-cell stage. (<i>lower panel)</i> Lysates of 12 hpf embryos were analyzed by immunoblotting for the presence of DeltaA. (higher panel) Graph quantifies 2 individual experiments, each with 30 injected embryos/group. (<b>B</b>), Fluorescent whole-mount antibody labeling of wild type (WT) and <i>asb11<sup>cul</sup></i> embryos at 24 hpf for the mitotic marker anti-phosphohistone-3 (PH 3) antibody (<i>green</i>) and the neuronal marker Hu(C). Graph shows the number of positive cells per area (5 somites from beginning of yolk extension) of 5 embryos for each genotype.</p
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