15 research outputs found

    Dex-IR inhibits the invasiveness of H1650 cells.

    No full text
    <p>(A) Invasion assay of H1650 cells treated with the drugs as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0194341#pone.0194341.g001" target="_blank">Fig 1</a>. The Transwell invasion assay showed that Dex-IR suppressed the invasion of H1650 cells. Images were captured at a magnification of 400× (A, left panel). Scale bars, 100 μm. Graphical representation of the number of invasive H1650 cells per microscopic field. Each column and bar shows the SEM from three independent experiments (*<i>P</i> < 0.05 <i>vs</i>. vehicle) (A, right panel). (B) Inhibition of MMP9 activity in conditioned medium from H1650 cells treated with Dex-IR at the indicated concentration and incubated for 18 h was evaluated using gelatin zymography. Representative data from a single experiment are shown. The left lanes are standard markers. (C) qRT-PCR analysis of the MMP2, MMP9, integrin α2, and integrin α5 gene expression in cells 6 h after treatment with drugs. Each experiment was repeated three times and the results shown are representative of the three independent experiments. The bar graph shows the mean ± SEM of three independent experiments (*P < 0.05 vs. MMP2 expression in vehicle; <sup>#</sup>P < 0.05 vs. MMP9 expression in vehicle; <sup>§</sup>P < 0.05 vs. integrin α2 expression in vehicle).</p

    Ionizing-radiation-irradiated Dex (Dex-IR) inhibits the proliferation of non-small cell lung cancer (NSCLC) cells.

    No full text
    <p>(A) The chromatograms of Dex (top) and the fraction of crude extracts with Dex-IR (bottom). The chromatography conditions are given in the Materials and Methods. The arrows indicate the retention time of each peak. (B) Lung cancer cell lines were treated with increasing concentrations of Dex and Dex-IR for 24 h. The effects of Dex-IR at the indicated concentration on the viability of lung cancer cells were determined using the MTT assay and were compared with those of Dex-treated cells. Data are presented as the mean ± standard error of the mean (SEM) of three independent experiments (*<i>P</i> < 0.05). (C) H1650 cells were treated with vehicle (1% DMSO), Dex (100 ug/mL), Dex-IR (100 ug/mL), or doxorubicin (DOXO; 1 μM) for 72 h. DOXO was used as a positive control. Cells were observed using phase-contrast microscopy (40× magnification). The scale bar is 200 μm.</p

    Increased apoptotic cell death induced by Dex-IR in H1650 lung cancer cells.

    No full text
    <p>(A) Annexin V/propidium iodide double staining analysis of apoptosis in H1650 cells. H1650 cells were treated with Dex, Dex-IR, or DOXO as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0194341#pone.0194341.g001" target="_blank">Fig 1</a> for 72 h. The bar graph shows the percentages of dead, living, early-apoptotic, and late-apoptotic cells according to treatment. Data are presented as the mean ± SEM of three independent experiments (*<i>P</i> < 0.05 <i>vs</i>. live cells in the medium; <sup>#</sup><i>P</i> < 0.05 <i>vs</i>. early apoptotic cells in the medium; and <sup>§</sup><i>P</i> < 0.05 <i>vs</i>. late apoptotic cells in the medium). (B) After treating H1650 cells with the same doses of the indicated drugs as described above for 24 h, the proteins were analyzed by Immunoblotting with antibodies against pro-Casp-3, cleaved Casp-3, PARP, and cleaved PARP. GAPDH was used to normalize the protein contents. (C) Dex-IR-induced apoptotic cells were detected by TUNEL assay (400× magnification). H1650 cells were treated with the drugs as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0194341#pone.0194341.g001" target="_blank">Fig 1</a> for 12 h.</p

    Dex-IR induced H1650 cell cycle arrest.

    No full text
    <p>(A) Cell cycle analysis following drug treatment for 72 h. The bar graph of the results of the cell cycle analysis of drug-treated H1650 cells shows the percentages of cells in different phases of the cell cycle (G<sub>0</sub>/G<sub>1</sub>, S, and G<sub>2</sub>/M). The bar graph shows the mean ± SEM of three independent experiments (*<i>P</i> < 0.05 <i>vs</i>. G<sub>0</sub>/G<sub>1</sub> phase in the medium; <sup>#</sup><i>P</i> < 0.05 <i>vs</i>. S phase in the medium; and <sup>§</sup><i>P</i> < 0.05 <i>vs</i>. G2/M phase in the medium). (B) Cell lysates were analyzed by Immunoblot analysis using specific antibodies against the cyclins A2, D1, B1 and Phospho-Rb. Protein loading was normalized based on GAPDH. (C) Cells were treated with 400 nM nocodazole for 24 h, and then further treated with Dex or Dex-IR for 24 h. They were then harvested, and subjected to cell cycle analysis. The bar graph shows the mean ± SEM of three independent experiments (*P < 0.05 vs. G<sub>0</sub>/G<sub>1</sub> phase in the Nocodazole).</p

    Delayed leaf senescence symptoms in the <i>ddm1-2</i> as well as in the <i>drd1-6</i> mutant.

    No full text
    <p>(A) Phenotypes of detached WT, <i>drd1-6</i>, <i>drd1-p</i> and <i>ddm1-2</i> leaves after 0, 3, and 5-d dark incubation. (B) Photochemical efficiency of photosystem II (Fv/Fm) in WT and mutant leaves in (A). Data represent average values ± SE (n = 27) of three independent experiments. Bars with the same letter are not significantly different at <i>P</i> < 0.05 by Tukey’s honestly significant difference (HSD) test.</p

    Delayed leaf senescence symptoms in the <i>drd1-6</i> mutant.

    No full text
    <p>(A) Phenotypes of 28-day-old and 55-day-old wild-type (WT) and <i>drd1-6</i> mutant whole plants. (B) Individually darkened leaf (IDL) senescence of WT (left) and <i>drd1-6</i> (right) plants. Rosette leaves of 28-day-old WT and <i>drd1-6</i> mutant (IDL 0 d) were induced to undergo senescence for 5 d under dark conditions (IDL 5 d). The red and blue arrows indicate 5 d IDL of WT and <i>drd1-6</i> plants, respectively. (C) Phenotypes of detached WT and <i>drd1-6</i> leaves after 5-d dark incubation. (D) Photochemical efficiency of photosystem II (Fv/Fm) and (E) maximal electron transport rate (ETRmax) in WT and the <i>drd1-6</i> leaves were examined at the indicated days during dark-induced senescence (DIS). Data represent average values ± SE (n = 27) of three independent experiments. * indicates <i>P</i> < 0.01 by student’s t-test.</p

    Transcript level changes in rosette leaves during DIS.

    No full text
    <p>(A) Microarray analysis represents that the numbers of genes show two- (black), three- (gray), or fourfold (white) up- (upper) or down-(lower) regulation in expression of 0 d, 3 d, and 5 d DIS <i>drd1-6</i> mutant compared to WT. (B) The number of genes with two-, three-, or fourfold increase or decrease in gene expression during 3 d and 5 d DIS compared to control (0 d) in the WT and the <i>drd1-6</i> mutant are represented by black, gray, and white bars, respectively. Data represent the means of two independent Affymetrix Gene Chip analyses. FD, fold difference.</p

    The positions of mutations and expression of <i>DRD1</i> gene.

    No full text
    <p>(A) Domain structures of DRD1 and DDM1 and positions of <i>drd1-6</i>, <i>drd1-p</i> and <i>ddm1-2</i>. Amino acid sequence change from tryptophan (W) to stop codon in helicase superfamily C-terminal (HELICc) domain in <i>drd1-6</i>. The triangle indicates the position of T-DNA insertion in <i>drd1-p</i> mutant. In case of <i>ddm1-2</i>, substitution of G to A in the splice donor site of intron 11 brings about lack of helicase superfamily C-terminal (HELICc) domain. (B) RT-PCR analysis of <i>DRD1</i> and control <i>ACTIN2</i> genes in WT and <i>drd1-p</i> mutant leaves. The <i>drd1-p</i> mutant displayed a decrease in <i>DRD1</i> expression levels compared to WT.</p

    Effect of CG extracts on liver toxicity using aspartate/alanine aminotransferase activity in Tg mice.

    No full text
    <p>The Mo/Hu APPswe PS1dE9 mice were fed with or without EA-CG (50 mg/kg BW) for 6 months. The brain were isolated from the EA-CG treated and control (PBS) groups (n = 3 mice per each group). After the brain was divided in half, we tested the gene expression profile of the inflammatory cytokines IL-1α (A) and IFN-γ (B) as markers of the cellular response by semi-quantitative RT-PCR. The fold change was presented after normalization to the housekeeping gene β-actin). *p < 0.05; n.s. mean not significant. The levels of Aspartate aminotransferase activity (AST, Units/L) and Alanine aminotransferase activity (ALT, Units/L)) were measured in the sera by centrifugation after obtain mice blood from each group, and then the relative value (U/L) of ALT (C) or AST (D) as hepatotoxicity markers were measured and presented after comparison to the controls (PBS groups), according to the manufacturer’s instructions. The values were presented as the means ± S.E.M (n = 3 mice per each group). n.s. mean not significant.</p

    Delayed leaf senescence symptoms in the <i>drd1-6</i> mutant at later developmental stages.

    No full text
    <p>Rosette leaves of 28-day-old WT and the 34, 36, and 38-day-old <i>drd1-6</i> mutants were detached and darkened for 0, 3, 5 days. Photochemical efficiency of photosystem II (Fv/Fm) in WT and mutant leaves was examined at the indicated days. Data represent average values ± SE (n = 20) of independent experiments. Bars with the same letter are not significantly different at <i>P</i> < 0.05 by Tukey’s honestly significant difference (HSD) test.</p
    corecore