Silencing of the reporter gene <i>URA5-miRs</i>.

<p>(A) The upper panels show a slower growth rate of the transformants of <i>URA5-miR1</i> (two transformants was picked in each case, namely, miR1-1 and miR1-2), and <i>URA5-miR2</i> (miR2-1 and miR2-2), than the wild type B4500, and the transformants of miR1-m (<i>i.e.</i> miR1-m1 and m2) and miR2-m, on MIN agar supplemented with 100 µg/ml hygromycin B, no hygromycin for B4500 and B4500FOA. The negative control B4500FOA (<i>ura5</i>) did not grow on MIN. On MINFOA (the bottom panels), the wild type strains and transformants of miR1-m and miR2-m were killed by FOA. Transformants containing miR1 and miR2, as well as <i>ura5</i>⁻ strain B4500FOA grew properly. MIN: minimal medium, and MINFOA: minimal medium with 5-FOA and 50 mg/l uracil. For selection of transformants, 100 µg/ml hygromycin B was added to MIN or MINFOA when needed. (B) Reverse transcription-PCR confirmed the decreased mRNA level of <i>URA5</i> in <i>URA5-miR</i> transformants. <i>URA5</i> mRNA levels in the wild type and in the transformants of miR-ms were close to each other. In each assay, two transformants were picked for double examination. (C) A semi-quantification of <i>URA5</i> mRNA to <i>ACT1</i> mRNA in the strains in (B).</p

miR-mediated gene silencing requires RNAi machinery in <i>C. neoformans</i>.

<p>(A) <i>URA5-miR1/2</i> restored the growth of B4500FOA (<i>ura5</i>) on MIN agar in the RNAi-deficient mutant strains, NE465, NE493, NE473 and NE475, <i>i.e.</i> gene silencing of <i>URA5-miRs</i> that was observed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052734#pone-0052734-g004" target="_blank">Fig. 4</a> did not occur in these mutants. And these transformants failed to grow on plates containing 5-FOA (the panels of MINFOA agar). MIN or MINFOA agar was supplemented with 100 µg/ml hygromycin B for the selection of all transformants of the reporters. The drug was left out for B4500 and B4500FOA. The transformants of miR1-m and miR2-m, together with the wild type B4500 and B4500FOA, served as control. (B) qRT-PCR confirmation of restored expression of <i>URA5</i> in RNAi-deficient mutants, NE465, NE493, NE473 and NE475, which are originally <i>ura5</i> defective strains.</p

Schematic diagram of the construction of reporters in silencing assay.

<p>The upper panel shows the construction of URA5-miRs or URA5-miR-ms. Two pair of primers, URA5-XhoI-S/URA5-miRs-BamHI and URA5-BamHI-S/URA5-XbaI-A, were used to PCR amplify the ORF and the terminator regions of <i>URA5</i>, respectively. Among the primers, URA5-miRs-BamHI harbored the sequence of miRs or miR-ms through primer design. The two PCR fragments were then ligated after digested by <i>BamH</i> I. Similarly, the construction of <i>CLC-miR1/2</i> and <i>CLC-miR1-m/miR2-m</i> was made (the bottom part). The position of miRs or miR-ms is indicated by the boxes. Restriction sites are in small letters. Arrows mark the position and direction of the primers. Start codons and stop codons of <i>URA5</i> and <i>CLC1</i> are underlined. For detailed description, see the section of Materials and Methods.</p

Carbon Consequences and Agricultural Implications of Growing Biofuel Crops on Marginal Agricultural Lands in China

Using marginal agricultural lands to grow energy crops for biofuel feedstocks is a promising option to meet the biofuel needs in populous China without causing further food shortages or environmental problems. Here we quantify the effects of growing switchgrass and Miscanthus on Chinese marginal agricultural lands on biomass production and carbon emissions with a global-scale biogeochemical model. We find that the national net primary production (NPP) of these two biofuel crops are 622 and 1546 g C m–2 yr–1, respectively, whereas the NPP of food crops is about 600 g C m–2 yr–1 in China. The net carbon sink over the 47 Mha of marginal agricultural lands across China is 2.1 Tg C yr–1 for switchgrass and 5.0 Tg C yr–1 for Miscanthus. Soil organic carbon is estimated to be 10 kg C m–2 in both biofuel ecosystems, which is equal to the soil carbon levels of grasslands in China. In order to reach the goal of 12.5 billion liters of bioethanol in 2020 using crop biomass as biofuel feedstocks, 7.9–8.0 Mha corn grain, 4.3–6.1 Mha switchgrass, or 1.4–2.0 Mha Miscanthus will be needed. Miscanthus has tremendous potential to meet future biofuel needs, and to benefit CO2 mitigation in China

Detection of miRNAs by Northern blotting.

<p>Positive bands for miR1 and miR2 (the left two panels) and their precursors were detected. On the right, rsRNA, a randomly picked small RNA only formed a smear band signal. Probes were the synthesized DNA sequence corresponding to miR1, miR2 or rsRNA. Total RNA was prepared from cultures collected at 18 hr and 36 hr as indicated. EtBr stands for ethium bromide-stained gel showing the 22-nt miRNA bands. Approximately 10 µg of total RNA was equally loaded. The size of the RNA markers is shown on left.</p

Expression of <i>CLC1-miRs</i> determined by qRT-PCR in different strains.

<p>Expression of <i>CLC1-miRs</i> determined by qRT-PCR in different strains.</p

Silencing of the reporter <i>CLC-miRs</i>.

<p>A similar silencing assay was carried out for the reporter <i>CLC-miRs</i>. (A) Melanin-deficient phenotype of the transformants of <i>CLC-miRs</i> (#1 to #3) was observed as expected, suggesting knockdown expression of <i>CLC1</i>. When miRs were mutated, <i>CLC-miR-ms</i> restored melanin production (Second and forth rows from the top). Cells were placed on low-glucose (0.1%) Asn agar with NE and 100 µg/ml hygromycin except for the wild types. (B) Decreased mRNA level of <i>CLC1</i> by miRs silencing determined by reverse transcription-PCR for the strains in (A). Relative abundance of <i>CLC1</i> mRNA verse actin-encoding gene <i>ACT1</i> mRNA was considered. PCR reaction was performed in triplicate.</p

Primers used in this study.

<p>Primers used in this study.</p

Additional file 7 of Comprehensive analysis of the expression, prognostic significance, and function of FAM83 family members in breast cancer

Additional file 7: Figure S7. Exploration the expression and subcellular localization of FAM83F by the HPA database

Blue Light-Excitable Broadband Yellow Emission in a Zero-Dimensional Hybrid Bismuth Halide with Type-II Band Alignment

Zero-dimensional (0D) organic–inorganic hybrid metal halides have captured broad interest in the lighting and display fields because of their unique electronic structures and splendid broadband emission properties. However, the blue light-excitable broadband yellow emissions have been rarely reported in 0D hybrid metal halides. Here, we design a new 0D bismuth hybrid, (4cmpyH)2BiCl5 (1, 4cmpy = 4-(chloromethyl)pyridine), featuring isolated edge-sharing bioctahedral [Bi2Cl10]4– dimers surrounded by rigid, conjugated, and luminescent organic [4cmpyH]+ cations. This material is able to show intrinsic broadband yellow emissions under blue light (468 nm) excitation with a long lifetime of 22.33 μs and a photoluminescence (PL) quantum yield of 5.56%. Solid-state UV–vis spectroscopy studies prove that introducing organic π-conjugated groups into hybrid systems leads to absorption in the visible light region, in favor of photoexcitation by visible light. By comparing the PL data of 1 and the organic template at room temperature and measuring variable-temperature PL spectra of 1, the blue light-excited broadband emission of 1 can be attributed to the synergistic emissions of intramolecular π → π* and n → π* transitions in the organic cations and triple self-trapped exciton (STE) states centralized at the highly distorted Bi–Cl lattices. Moreover, density functional theory calculations reveal a type-II band alignment in 1 with an indirect band gap of 2.64 eV, which is together determined by organic cations and inorganic bioctahedral units. To the best of our knowledge, our work represents the first report on the blue light-excitable STE emission in 0D Bi-based metal halides, which will largely promote the rapid development of novel high-performance yellow light-emitting materials