Data of Lake Gonghai

Data of climate proxies from Lake Gonghai since last deglaciation to 7 ka BP. Including pollen-based precipitation reconstruction; % tree pollen; % herb pollen; organic matter content; magnetic susceptibility; mean grain size; carbonate content. (Original reference: Chen, F. H., Xu, Q. H., Chen, J. H., Birks, H. J., Liu, J. B., Zhang, S. R., et al. (2015). East Asian summer monsoon precipitation variability since the last deglaciation. <i>Scientific Reports</i>, 5, 11186.

sv40-miR-S1-5p and hsa-miR423-5p downregulate the predicted biological targets of hsa-miR423-5p.

<p>HeLa cultures were transfected by the indicator vectors pmiR-S1-5p:DMWDUTR, pmiR423-5p:DMWDUTR, pmiR-S1-5p:C20orf27UTR, pmiR423-5p:C20orf27UTR and their control vectors with miRNAs only, with miRNA deletions or without miRNA pairing sites in the 3′-UTR.</p

The construction and principle behind the dual-fluorescent protein reporter vector.

<p>(A) A diagram of the dual-fluorescent protein reporter vector pMGhU6. (B) The principle of the dual-fluorescent protein reporter vector system.</p

Validation of the dual-fluorescent protein reporter system as an miRNA functional assay.

<p>(A) Sequences of miR30, hiv1-miR-N367 and their non-fully complementary targets. (B) Northern blot analysis of the transcription of specific miRNAs. HeLa cells were mock-transfected (mock) or transfected with plasmids pmiR30, pmiR-N367 individually, and the location of the mature miR30 and miR-N367 is indicated. tRNA^Val served as a loading control <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036157#pone.0036157-Tran1" target="_blank">[26]</a>. (C) Fluorescence reporter assay of the indicator vectors pmiR30:4tar(30), pmiR-N367:4tar(n) and their control vectors. HeLa cell cultures were transfected with the indicator vectors pmiR30:4tar(30), pmiR-N367:4tar(n) and their control vectors with miRNAs or target sequences only. The ratio of EGFP to mCherry fluorescence intensity is shown.</p

hiv1-miR-N367 and hsa-miR192 act as functional orthologs.

<p>(A) Sequence homology between hiv1-miR-N367 and hsa-miR192 (marked in red). The seed regions of miRNAs were underlined. (B) Sequences of miR192 and its non-fully complementary target. (C) Fluorescence reporter assay of the indicator vectors pmiR-N367:4tar(n), pmiR192:4tar(n), pmiR-N367:4tar(192), pmiR192:4tar(192) and their control vectors with miRNAs or target sequences only. (D) Fluorescence reporter assay of the indicator vectors pmiR-N367:PABPC4UTR, pmiR192:PABPC4UTR and their control vectors with miRNAs only, with miRNA deletions or without miRNA pairing sites in the 3′-UTR.</p

Monitoring miRNA activities in living cells.

<p>Activities of two miRNAs, including miR30 (A), miR-N367 (B), were imaged in live HeLa cell cultures that were transfected with the indicator vectors pmiR30:4tar(30), pmiR-N367:4tar(n) and their control vectors with miRNAs or target sequences only, respectively. Bar, 10 μm.</p

Sub-ppt Mass Spectrometric Detection of Therapeutic Drugs in Complex Biological Matrixes Using Polystyrene-Microsphere-Coated Paper Spray

Polystyrene (PS) is a class of polymer materials that offers great potential for various applications. However, the applications of PS microspheres in paper spray mass spectrometry are largely underexplored. Herein we prepared a series of PS microspheres via a simple dispersion polymerization and then used them as coating materials for paper spray mass spectrometry (MS) in high-sensitivity analysis of various therapeutic drugs in complex biological matrixes. In the preparation of PS-coated papers, the coating method was found playing a key role in determining the performance of the resulting paper substrate in addition to other parameters (e.g., starch type and amount, PS coating amount, and spray solvent). We also found that as a solvent was applied on PS-coated paper for paper spray, the analytes of interest would be first extracted out and then moved to the tip of paper triangle for spray along with the applied solvent. In the process, the surface energy of PS particles had a strong impact on the desorption performance of analytes from PS-coated paper substrate, and the PS with a high surface energy favored the elution of analytes to allow a high MS sensitivity. When the prepared PS coated paper was used as a substrate for paper spray, it gave high sensitivity in analysis of therapeutic drugs in various biological matrixes such as whole blood, serum, and urine with excellent repeatability and reproducibility. In contrast to uncoated filter paper, an improvement of 10–546-fold in sensitivity was achieved using PS-coated paper for paper spray, and an estimated lower limit of quantitation (LLOQs) in the range of 0.004–0.084 ng mL^–1 was obtained. The present study is significant in exploring the potential of PS for high-sensitivity MS analysis, and it provides a promising platform in the translation of the MS technique to clinical applications

Three-Fragment Fluorescence Complementation Coupled with Photoactivated Localization Microscopy for Nanoscale Imaging of Ternary Complexes

Many cellular processes are governed by molecular machineries that involve multiple protein interactions. However, visualizing and identifying multiprotein complexes such as ternary complexes inside cells is always challenging, particularly in the subdiffraction cellular space. Here, we developed a three-fragment fluorescence complementation system (TFFC) based on the splitting of a photoactivatable fluorescent protein, mIrisFP, for the imaging of ternary complexes inside living cells. Using a combination of TFFC and photoactivated localization microscopy (PALM), namely, the TFFC-PALM technique, we are able to identify the multi-interaction of a ternary complex with nanometer-level spatial resolution and single-molecule sensitivity. The TFFC-PALM system has been further applied to the analysis of the G_s ternary complex, which is composed of α_s, β₁, and γ₂ subunits, providing further insights into the subcellular localization and function of G protein subunits at the single-molecule level. The TFFC-PALM represents a valuable method for the visualization and identification of ternary complexes inside cells at the nanometer scale

In Vivo Targeting and Imaging of Atherosclerosis Using Multifunctional Virus-Like Particles of Simian Virus 40

Atherosclerosis is a leading cause of death globally. Targeted imaging and therapeutics are desirable for the detection and treatment of the disease. In this study, we developed trifunctional Simian virus 40 (SV40)-based nanoparticles for in vivo targeting and imaging of atherosclerotic plaques. These novel trifunctional SV40-based nanoparticles encapsulate near-infrared quantum dots and bear a targeting element and a drug component. Using trifunctional SV40-based nanoparticles, we were able to noninvasively fluorescently image atherosclerotic plaques in live intact ApoE­(−/−) mice. Near-infrared quantum dots encapsulated in the SV40 virus-like particles showed prominent optical properties for in vivo imaging. When different targeting peptides for vascular cell adhesion molecule-1, macrophages, and fibrin were used, early, developmental, and late stages of atherosclerosis could be targeted and imaged in live intact ApoE­(−/−) mice, respectively. Targeted SV40 virus-like particles also delivered an increased concentration of the anticoagulant drug Hirulog to atherosclerosis plaques. Our study provides novel SV40-based nanoparticles with multivalency and multifunctionality suitable for in vivo imaging, molecular targeting, and drug delivery in atherosclerosis

Delaying Photobleaching of a Light-Switch Complex for Real-Time Imaging of Single Viral Particle Uncoating

Photobleaching is a major obstacle in the real-time imaging of biological events, particularly at the single-molecule/particle level. Here, we report a strategy to delay photobleaching of a light-switch complex, [Ru­(phen)₂dppx]²⁺, by insertion of a six-cysteine peptide into virus particles. The six-cysteine peptide was inserted into viral protein R of HIV-1 and assembled into infectious HIV-1 viral particles, where it effectively delayed the photobleaching of the [Ru­(phen)₂dppx]²⁺ complex used to label viral genomic RNAs. This delay in photobleaching allowed for a monofluorescent assay to be constructed for the real-time monitoring of viral uncoating, a poorly understood process. This novel strategy to delay photobleaching in infectious viral particles provides a powerful method to analyze viral uncoating at the single-particle level in real time