Membrane binding analyses of CrtQ-FLAG and CrtO-FLAG.

<p>Western blots of soluble fractions vs. total membranes (5 μg each) isolated from both strains and the effect of different washes on CrtO and CrtQ membrane association. Total membranes were washed with EB8, EB12 or EB8+U and soluble (“Sol.”) and pellet (“Pel.”) fractions were generated by ultracentrifugation. Each lane was loaded with equal volumes of (resuspended) pellet or supernatant fractions. Three independent cultures were tested in this manner. (A). Western blots analysis of anti-FLAG resin eluates from 2% DDM-solubilized total membranes of WT, CrtO-FLAG and CrtQ-FLAG cells, separated on 4–18% Clear Native–PAGE (B). Colour scan of CN-PAGE gel for WT total membrane showing the relative migration of several known complexes.</p

Carotenoid distribution in <i>Synechocystis</i> membranes.

<p>Total carotenoid amount in purified PM1, PM2 and TM (A). Relative amount of different carotenoids in each membrane (B). Data represent means ± standard deviations for carotenoids in membranes from three independent cultures. For results of statistical analysis, please refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130904#pone.0130904.s006" target="_blank">S2 Table</a>.</p

A simplified pathway for carotenoid biosynthesis in <i>Synehocystis</i>.

<p>Only enzymes with verified function are included.</p

Effect of Different Groundwater Levels on Seismic Dynamic Response and Failure Mode of Sandy Slope

<div><p>Heavy seismic damage tends to occur in slopes when groundwater is present. The main objectives of this paper are to determine the dynamic response and failure mode of sandy slope subjected simultaneously to seismic forces and variable groundwater conditions. This paper applies the finite element method, which is a fast and efficient design tool in modern engineering analysis, to evaluate dynamic response of the slope subjected simultaneously to seismic forces and variable groundwater conditions. Shaking table test is conducted to analyze the failure mode and verify the accuracy of the finite element method results. The research results show that dynamic response values of the slope have different variation rules under near and far field earthquakes. And the damage location and pattern of the slope are different in varying groundwater conditions. The destruction starts at the top of the slope when the slope is in no groundwater, which shows that the slope appears obvious whipping effect under the earthquake. The destruction starts at the toe of the slope when the slope is in the high groundwater levels. Meanwhile, the top of the slope shows obvious seismic subsidence phenomenon after earthquake. Furthermore, the existence of the groundwater has a certain effect of damping.</p></div

CrtQ and CrtO localization.

<p>Western blots analyses of purified PM1, PM2 and TM from CrtQ-FLAG and CrtO-FLAG strains. Exposure times were 4 and 180 seconds respectively (A). Quantifications of CrtQ (grey) and CrtO (white) in each membrane, based on four independent western blots (B). Data represent means ± standard deviations for four independent quantifications. For results of statistical analysis, please refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130904#pone.0130904.s006" target="_blank">S2 Table</a>.</p

Basic earthquake acceleration value.

<p>Basic earthquake acceleration value.</p

Intron Definition and a Branch Site Adenosine at nt 385 Control RNA Splicing of HPV16 E6*I and E7 Expression

<div><p>HPV16 E6 and E7, two viral oncogenes, are expressed from a single bicistronic pre-mRNA. In this report, we provide the evidence that the bicistronic pre-mRNA intron 1 contains three 5′ splice sites (5′ ss) and three 3′ splice sites (3′ ss) normally used in HPV16⁺ cervical cancer and its derived cell lines. The choice of two novel alternative 5′ ss (nt 221 5′ ss and nt 191 5′ ss) produces two novel isoforms of E6E7 mRNAs (E6*V and E6*VI). The nt 226 5′ ss and nt 409 3′ ss is preferentially selected over the other splice sites crossing over the intron to excise a minimal length of the intron in RNA splicing. We identified AACAAAC as the preferred branch point sequence (BPS) and an adenosine at nt 385 (underlined) in the BPS as a branch site to dictate the selection of the nt 409 3′ ss for E6*I splicing and E7 expression. Introduction of point mutations into the mapped BPS led to reduced U2 binding to the BPS and thereby inhibition of the second step of E6E7 splicing at the nt 409 3′ ss. Importantly, the E6E7 bicistronic RNA with a mutant BPS and inefficient splicing makes little or no E7 and the resulted E6 with mutations of ⁹¹QYNK⁹⁴ to ⁹¹PSFW⁹⁴ displays attenuate activity on p53 degradation. Together, our data provide structural basis of the E6E7 intron 1 for better understanding of how viral E6 and E7 expression is regulated by alternative RNA splicing. This study elucidates for the first time a mapped branch point in HPV16 genome involved in viral oncogene expression.</p> </div

Similarity constants.

<p>Similarity constants.</p

Cluster miRNA targets expression profile.

<p>A: Model of miRNA target expression pattern. B: Heatmap of miRNA target expression profile. The cluster was done on the basis of log₂ (expression level in treatment/expression level in control). Yellow shows down-regulation. Blue shows up-regulation.</p

Expression of zma-miR528a/b and one of its targets.

<p>A: SLRT-PCR results of miRNA528a/b at 3 biological replicates. B: qRT-PCR results of one of miRNA528a/b targets GRMZM2G039381 at 3 biological replicates. X-axis shows the inbred lines under different time point treatment compared to control. Y-axis shows the expression level. Error bars show the standard deviation. Student t test was performed on qRT-PCR results. ** denotes the p value <0.01 and * denotes the p value <0.05.</p