Specificity of pAb506P.

<p><b>(A)</b> Comparative titration curves of pAb506P on ELISA plates coated with a topo I peptide surrounding the serine 506 site, either in its phosphorylated or non-phosphorylated form. <b>(B)</b> Western analyses of H358 cell lysates (100 μg/lane) probed with pAb506P (lane 1) or with goat anti-topo I (lane 2). Arrows indicate positions of the 45 kDa species and full length topo I. <b>(C)</b> Topo I immunoprecipitation (goat anti-topo I C-terminus) followed by pAb506P Western of H358 cell lysates. Lane represents 200 μg starting material. <b>(D)</b>Western analysis of PS506 and actin in H358 cell lysates before (cntr) and after treatment with alkaline phosphatase (AP). <b>(E)</b> Western analysis of PS506 (using pAb506P), full length topo I (using goat anti-topo I) and tubulin in H358 cells before and after a 24 hr treatment of cells with 0.1 or 1 μM CPT.</p

Expression of PS506 in malignant, non-malignant, and benign tumor tissues.

<p><b>(A)</b> Representative PS506 Western blot of specimens of NSCLC and their paired non-malignant specimens (specimen pairs 8–13). H358 is the reference control for quantification of all PS506 blots. Tables <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134929#pone.0134929.t001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134929#pone.0134929.t002" target="_blank">2</a> list the specimens analyzed and quantification of PS506 levels relative to H358. <b>(B)</b> Bar graph of the PS506 levels shown in Tables <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134929#pone.0134929.t001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134929#pone.0134929.t002" target="_blank">2</a>, grouped as paired malignant/non-malignant specimens and benign tumors. <b>(C)</b> Scatter plot of PS506 levels from Tables <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134929#pone.0134929.t001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134929#pone.0134929.t002" target="_blank">2</a>, grouped as malignant (M), non-malignant (N), and benign (B) tumors. The p values were calculated by an unpaired t-test.</p

Data File 1: Design of ultracompact polarimeters based on dielectric metasurfaces

Design data for a x-polarized sensitive flat focusing lens. Originally published in Optics Letters on 15 April 2017 (ol-42-8-1580

Demonstration of Polymorphic Spacing Strategy against Sintering: Synthesis of Stabilized Calcium Looping Absorbents for High-Temperature CO₂ Sorption

To decrease the sintering deterioration of CaO sorbents in multiple CO₂ capture and release cycles, we synthesized a series of stabilized CaO sorbents incorporated with silica through freeze-drying and heat-drying, the latter of which was referred to as benchmark. The ratio of Ca and Si precursors was varied to control the reactive loadings of CaO (from 70 to 100 wt %) and the fraction of spacers in the sorbents. The characterization results show that the freeze-drying method produces sorbents with higher specific area and larger pore volume than the heat-drying method. Moreover, the stability test of over 30 cycles demonstrated that the freeze-dried samples exhibited better performance with higher stability and total CO₂ uptake. The high-resolution transmission electron microscopy image shows that Ca₂SiO₄ crystallites as spacers are distributed within the matrix of CaO crystallites. The optimal spacer loading was determined to be ∼10 wt %, and the optimal reaction temperature was found to be 700 °C. Finally, the best sorbent was tested under harsh conditions and maintained a stable capture capacity with a CO₂ uptake of 0.21 g of CO₂ g^–1 of sorbent even at the 30th cycle. The performance of the sorbent in this work was then systematically compared to those reported in the literature. The use of a Si-based spacer and freeze-drying have significant potential to enhance the stability of CaO sorbents

Novel Role of Src in Priming Pyk2 Phosphorylation

<div><p>Proline-rich tyrosine kinase 2 (Pyk2) is a member of the focal adhesion kinase (FAK) family of non-receptor tyrosine kinases and plays an important role in diverse cellular events downstream of the integrin-family of receptors, including cell migration, proliferation and survival. Here, we have identified a novel role for Src kinase in priming Pyk2 phosphorylation and subsequent activation upon cell attachment on the integrin-ligand fibronectin. By using complementary methods, we show that Src activity is indispensable for the initial Pyk2 phosphorylation on the Y402 site observed in response to cell attachment. In contrast, the initial fibronectin-induced autophosphorylation of FAK in the homologous Y397 site occurs in a Src-independent manner. We demonstrate that the SH2-domain of Src is required for Src binding to Pyk2 and for Pyk2 phosphorylation at sites Y402 and Y579. Moreover, Y402 phosphorylation is a prerequisite for the subsequent Y579 phosphorylation. While this initial phosphorylation of Pyk2 by Src is independent of Pyk2 kinase activity, subsequent autophosphorylation of Pyk2 <i>in trans</i> is required for full Pyk2 phosphorylation and activation. Collectively, our studies reveal a novel function of Src in priming Pyk2 (but not FAK) phosphorylation and subsequent activation downstream of integrins, and shed light on the signaling events that regulate the function of Pyk2.</p></div

Schematic representation of Pyk2 phosphorylation triggered by Src.

<p>In the inactive state, Pyk2 is in an unphosphorylated state. Upon integrin ligand binding, Src phosphorylates Y402, creating a binding site for the SH2-domain of Src, and enabling further phosphorylation of Pyk2 by Src, including at site Y579 (and likely at Y580, although not studied here). As shown by others [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0149231#pone.0149231.ref021" target="_blank">21</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0149231#pone.0149231.ref022" target="_blank">22</a>] phosphorylated and activated Pyk2 dimerizes or oligomerizes with itself, leading to further phosphorylation of Pyk2 via auto/trans-phosphorylation.</p

Detailed Investigations of the Countercurrent Multiphase (Gas–Liquid and Gas–Liquid–Liquid) Flow Behavior by Three-Dimensional Computational Fluid Dynamics Simulations

This paper focuses on the detailed investigations of the (multi-) liquid flow behaviors on an inclined stainless steel plate in simulations and experiments. Simulations are carried out with FLUENT 6.3; a three-dimensional computational fluid dynamics model considering the gravity, surface tension, and local drag force is developed and presented. Experimental data is obtained with optical methods. One main aspect of the paper’s presentation is the validation of the developed model for the case of one-liquid phase with a countercurrent gas phase, where the parameters (e.g., the thickness and the velocity profile of the liquid phase) are compared with the simulation results and the experimental data. The other key part of the presentation is the numerical investigations with the developed model for the case of two immiscible liquid phases with a countercurrent gas phase. Results show that with the current gas flow rate, the complex flow behavior strongly depends on the (multi-) liquid flow rates and the feed sequence. Furthermore, there is a high stability between the two liquids. All of these data lead to a better understanding of the complex fluid dynamics in three-phase distillation processes

Src activity is required for Pyk2 phosphorylation in cells plated on fibronectin.

<p>A, serum-starved HeLa cells were detached and kept in suspension in DMEM containing 0.5% BSA for 1 hr. Cells were then seeded on fibronectin- or poly-l-lysine (PLL)-coated dishes, and allowed to attach for the indicated times. Phosphorylation of Pyk2 and FAK was examined by immunoblot, as indicated. B, HeLa cells were kept in suspension with or without indicated Src inhibitors for 1 hr before the attachment to fibronectin-coated dishes for 15 min, and phosphorylation of Pyk2 and FAK was examined as above. C, HeLa cells transfected with mock, wild-type Src (Src-WT) or kinase-dead mutant (Src-KD) were kept in suspension for 1 hr then were attached to fibronectin-coated dishes for 15 min, and the phosphorylation of Pyk2 and FAK was examined by immunoblot as indicated. D, serum-starved SYF and Src⁺⁺ MEFs were kept in suspension for 1 hr, then were attached to fibronectin-coated dishes for 15 or 30 min as indicated, and the phosphorylation of Pyk2 and FAK was examined as in above.</p

Novel CaO–SiO₂ Sorbent and Bifunctional Ni/Co–CaO/SiO₂ Complex for Selective H₂ Synthesis from Cellulose

Catalysis- and sorption-enhanced biomass gasification is a promising route to high-purity hydrogen (H₂); however, most CaO-based sorbents for CO₂ capture have poor surface area and mechanical properties, lose carrying capacity over multiple uses, and have insufficient porosity to accommodate extra catalyst sites. We aimed to develop a high-surface-area CaO–SiO₂ framework onto which catalysts could be grafted. The best CaO–SiO₂ sorbent (<i>n</i>_Ca/<i>n</i>_Si = 2:1) maintained a CaO conversion of 65% even after 50 carbonation–decarbonation cycles, better than commercial micrometer-sized CaO or tailored CaO, because of stabilization via Ca–O–Si interactions and an ordered porous structure. Bimetallic catalyst grains (Ni/Co alloy, <20 nm) could be evenly loaded onto this structure by impregnation. The resulting bifunctional complex produced H₂ at nearly the same rate as a mixture of catalyst and commercial CaO while using less total sorbent/catalyst. Furthermore, this complex was much more durable due to its higher coking resistance and stable structure. After 25 carbonation–decarbonation cycles, the new catalyst–sorbent complex enhanced the H₂ yield from cellulose far more than a mixture of catalyst and commercial CaO did following the same treatment

Src is required to initiate adhesion-induced Pyk2 phosphorylation.

<p>A, 293T cells were transfected with GFP-tagged Pyk2-WT, Pyk2-Y402F or Pyk2-KD, with or without Src, serum-starved, detached, and then reattached to fibronectin-coated dishes for 30 min. Cell lysates were collected and assayed by immunoblot to examine Pyk2 phosphorylation. B, SYF cells were transfected and assayed similarly as in A. C, SYF cells were co-transfected with Src and GFP-tagged Pyk2-WT, or Pyk2-Y402F, or Pyk2-KD as indicated. The expressed Src was immunoprecipitated, and the associated Pyk2 was examined by immunoblot. D, GST-Pyk2-WT and GST-Pyk2-Y402F were expressed in <i>E</i>. <i>coli</i>, purified, and employed in an in vitro kinase assay as described in Materials and Methods. Pyk2 phosphorylation by recombinant Src was examined by immunoblot analysis by using anti-phospho-Pyk2 (Y402) antibody.</p