14,274 research outputs found
RT-PCR analysis of mRNA levels of ATG8 family members in ATG8f silenced plants.
No full textThe semi-quantitative RT-PCR analysis was conducted on the same set of plant samples to assess the effectiveness of ATG8f silencing. The second panel shows comparable mRNA levels of ATG8a and bottom panel for ATG8i, indicating selective gene silencing of ATG8f. Top panel: The RT-PCR analysis of tubulin mRNA level in the ATG8f-silenced (lanes 4–6) and control (lanes 1–3) plants. The panels were from the same gels, respectively. The experiment was repeated. (TIF)</p
Morpholino-mediated knockdown of <i>lin-28a</i> and <i>lin-28b</i> affects early development.
No full text<p>A. B. Schematic representation shows the genomic organization of the <i>lin-28a</i> and <i>lin-28b</i> genes in zebrafish. Regions targeted by splice-blocking morpholinos are shown. Arrows depict the location of the two primer sets to amplify exon and intron sequences, which were utilized in the RT-PCR analysis. C. D. The efficacy of MO was validated by RT-PCR using primers as indicated in A, B. Total RNA was isolated from <i>lin-28a</i>, <i>lin-28b</i> or control MO-injected embryos at 5 hpf and analyzed by using specific primers for <i>lin-28a</i> and <i>lin-28b</i>. Upper panel shows a reduced expression of <i>lin-28a</i> and <i>lin-28b mRNA</i>, and an increase of an aberrantly spliced <i>lin-28b</i> transcripts corresponding to the presence of intron 1. Lower panel shows RT-PCR products with <i>gapdh</i> primers used as internal control. E. Representative images of severe phenotype of <i>lin-28a</i> and <i>lin-28b</i> morphants at 5 hpf. F. Vegetal view of control, <i>lin-28a</i> and <i>lin-28b</i> morphants at 8 hpf. Arrow shows the similar extent of prechordal plate extension in control, <i>lin-28a</i> and <i>lin-28b</i> morphant embryos; bracket shows a reduced extent in epiboly in <i>lin-28a</i> and <i>lin-28b</i> morphant embryos compared with controls. G. Representatives of mild and severe phenotypes in <i>lin-28a</i> and <i>lin-28b</i> morphants at 30 hpf. Both <i>lin-28a</i> and <i>lin-28b</i> morphants develop morphological phenotypes displaying shorten body axis, small anterior structures and aberrant tail morphology ranging from mild (Right) to severe (Left) compare to control MO injected embryos (Top). Arrowheads indicate the smaller head. H. Quantification of the efficiency of rescue from gastrulation defects following co-injection of <i>lin-28a</i> MO, <i>lin-28b</i> MO and mRNAs. More than half of the <i>lin-28a</i> MO or <i>lin-28b</i> MO injected embryos had died by 30 hpf. The frequencies of the phenotypes were similar between <i>lin-28a</i> and <i>lin-28b</i> morphants. The frequency of the dead, mild and severe phenotypes decreased with a rescue experiment in which 300 pg of <i>lin-28a</i> and <i>lin-28b</i> mRNAs were co-injected with the MO. (p<0.0001, <i>lin-28a</i> MO vs. <i>lin-28a</i> MO+<i>lin-28a</i> mRNA, <i>lin-28b</i> MO vs. <i>lin-28b</i> MO+<i>lin-28b</i> mRNA, Fisher’s exact test). The total number of embryos is noted above each bar.</p
Vascular endothelial growth factor restores delayed tumor progression in tumors depleted of macrophages
No full textGenetic depletion of macrophages in Polyoma Middle T oncoprotein (PyMT)-induced mammary tumors in mice delayed the angiogenic switch and the progression to malignancy. To determine whether vascular endothelial growth factor A (VEGF-A) produced by tumor-associated macrophages regulated the onset of the angiogenic switch, a genetic approach was used to restore expression of VEGF-A into tumors at the benign stages. This stimulated formation of a high-density vessel network and in macrophage-depleted mice, was followed by accelerated tumor progression. The expression of VEGF-A led to a massive infiltration into the tumor of leukocytes that were mostly macrophages. This study suggests that macrophage-produced VEGF regulates malignant progression through stimulating tumor angiogenesis, leukocytic infiltration and tumor cell invasion
The relationship between co-authorship, currency of references and author self-citations
No full textThis paper attempts to identify the relationship between co-authorship and the currency of the references and author self-citations in the key journals of environmental engineering. The results show that the self-citation rate of co-authored articles is higher than in single-authored articles. A statistically significant correlation is identified between the numbers of co-authors, the rate of author self-citing and the author self-cited; though it was a low correlation. The value of coefficient correlation between the number of co-authors and the author self-citing rate is slightly higher than that between the number of co-authors and the author self-cited rate, which indicates that the number of co-authors hold a stronger correlation with the self-citing rate than the self-cited rate. Meanwhile, self-citing references are found to be more up-to-date than references to others. The range of publication years of self-citing references is smaller than that of references to others, indicating that researchers tend to preferentially cite their own recent works. There is no significant difference in the latest references between self-citing references and the references to others. It might result from electronic journals that provide an easy access to the most current publications.補正完畢國外SSCIY紙本電子版HU
Effects of adding nano metal powers on thermooxidative degradation of poly(Ethylene Glycol).
No full textZ. Lin, X. Han, T. Wang and S. L
RT-qPCR analyses relative transcript levels of <i>gntR</i> in WT SS2 at different growth stages.
No full textRT-qPCR analyses relative transcript levels of gntR in WT SS2 at different growth stages.</p
Expression of miRNAs and downstream heterochronic genes in <i>lin-28a</i> and <i>lin-28b</i> morphant embryos.
No full text<p>A. miRNA expression profiling in <i>lin-28a</i> and <i>lin-28b</i> morphants embryos at 5 hpf. Hierarchically clustered heat-map representing differences in miRNA expression between control MO-injected embryos and <i>lin-28a</i> MO- or <i>lin-28b</i> MO-injected embryos. Relative levels of expression are colored from green (low) to red (high). B. Real-time PCR analysis of <i>let-7a, let-7b, miR-430a</i> and <i>miR-430b</i> expression in control MO-, <i>lin-28a</i> MO- and <i>lin-28b</i> MO-injected embryos. Expression levels of <i>let-7a, let-7b, miR-430a</i> and <i>miR-430b</i> were analyzed at 5 hpf using TaqMan miRNA assay. The expression level of <i>miR-430a</i> and <i>miR-430b</i> was downregulated in both <i>lin-28a</i> MO- and <i>lin-28b</i> MO-injected embryos compared with control MO-injected embryos (***p<0.001, control MO vs. <i>lin-28a</i> MO, control MO vs. <i>lin-28b</i> MO, two-tailed test, n = 3, mean ± SEM). C. Real-time PCR analysis of <i>let-7a</i> and <i>let-7b</i> expression in control MO-, lin-28a MO- and lin-28b MO-injected embryos. Expression levels of <i>let-7a</i> and <i>let-7b</i> were analyzed using TaqMan miRNA assay at 28 hpf. The expression level of <i>let-7a</i> and <i>let-7b</i> was increased in both <i>lin-28a</i> MO- and <i>lin-28b</i> MO-injected embryos embryos compared with control MO-injected embryos (*p<0.05, **p<0.01, control MO vs. <i>lin-28a</i> MO, control MO vs. <i>lin-28b</i> MO, two-tailed test, n = 4, mean ± SEM). D. Semiquantitative RT-PCR analysis of <i>lin-41</i> gene expression in control MO-, lin-28a MO- and lin-28b MO-injected embryos. Total RNA was isolated from <i>lin-28a</i> MO-, <i>lin-28b</i> MO- or control MO-injected embryos at 24 hpf and analyzed by using specific primers for <i>lin-41.</i> The expression of <i>lin-41</i> was decreased in both <i>lin-28a</i> MO- and <i>lin-28b</i> MO-injected embryos compared with control MO-injected embryos. E. Control, <i>lin-28a</i> and <i>lin-28b</i> MO-injected embryos labeled by <i>in situ</i> hybridization with <i>lin-41</i> probe. Expression of <i>lin-41</i> was repressed in CNS of both <i>lin-28a</i> and <i>lin-28b</i> morphant embryos at 24 hpf.</p
The lin-4 Gene Controls Fat Accumulation and Longevity in Caenorhabditis elegans
No full textPrevious studies have determined that lin-4, which was the first miRNA to be discovered, controls the timing of cell fate determination and life span in Caenorhabditis elegans. However, the mechanism of lin-4 involvement in these processes remains poorly understood. Fat storage is an essential aspect of the life cycle of organisms, and the function of lin-4 in fat accumulation is not clear. In this study, we showed that the fat content is reduced remarkably in C. elegans lin-4 mutants. Quantitative RT-PCR analysis revealed a considerable decrease in the levels of SBP-1 and OGA-1 mRNA in lin-4 mutants. We also showed that lin-4 mutants have a significantly shorter life span than wild-type worms. DCF assay experiments showed that the reactive oxygen species (ROS) levels increased and mitochondrial DNA (mtDNA) copy number decreased in loss-of-function lin-4 mutants. These mutants also showed attenuation of locomotion. Taken together, our findings suggest that lin-4 may play an important role in regulating fat accumulation and locomotion and that lin-4 may control the life span of C. elegans by mediating ROS production
Freshly isolated Lin-SP cells express FAPs surface markers but are capable of myogenic differentiation.
No full text<p><b>A</b>: RT-PCR analysis of freshly isolated Lin- SP cells from wild type (WT) and <i>mdx<sup>5cv</sup></i> (MDX) muscle for myogenic markers (Pax7 and Myf5) and FAPs markers (PDGFRα and Sca1). Positive controls (PC) are sorted Sca1-positive cells for Sca1 and Lin- MP cells for Pax7, Myf5 and PDGFRα. Negative controls (NC) are sorted Sca1-negative cells for Sca1 and CD45-positive MP cells for Pax7, Myf5 and PDGFRα. <b>B, C</b>: FACS analysis of PDGFRα and Sca1 protein expression in Lin- SP cells (<b>B</b>) and Lin- MP cells (<b>C</b>) from wild type (WT) and <i>mdx<sup>5cv</sup></i> (MDX) muscle. Percentages of cells double positive (red) and double negative (green) for PDGFRα and Sca1 are shown. <b>D</b>: Confirmation by RT-PCR for PDGFRα expression in Lin- SP and Lin- MP cells sorted into PDGFRα-positive (Pα+) and PDGFRα-negative (Pα−) sub-fractions. <b>E</b>: <i>In vitro</i> myogenic differentiation of Lin- SP and MP cells sorted based on PDGFRα (Pα) expression. Cells were fixed after 14 days in culture and immunolabelled for α-actinin (green) to reveal myotubes. Cultures were counterstained with DAPI (blue) to visualize nuclei. Lin- MP Pα+ cells correspond to the previously characterized FAPs. Lin- MP Pα− cells are enriched in myogenic cells and also contain fibroblasts. Cultured wild type Lin- MP Pα− cells had 2,261 myotubes while cultured <i>mdx<sup>5cv</sup></i> Lin- MP Pα− cells had only 541 myotubes. Lin- SP Pα− cells did not survive in culture and are not shown. Scale bar = 400 μm. <b>E</b>: Cultures of Lin- SP Pα− cells were double labeled with antibodies to α-actinin (green) and collagen I (red) to visualize myotubes and fibroblasts, respectively. Cultures of Lin- SP Pα− cells from dystrophic muscle (MDX) do not contain myotubes but give rise to fibroblasts. Scale bar = 100 μm. <b>G</b>. Quantitative RT-PCR for Pax7 expression in Pdgfrα+ and Pdgfrα− Lin-SP cells, and Pdgfrα− Lin-MP cells. Data is presented as means +/− s.d. from 3 technical replicates.</p
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