7 research outputs found

    <i>Aopub1</i> disruptant showed defects in the conidia formation and more severe growth retardation in stress conditions.

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    <p>(A) Stress-induced formation of stress granules (indicated by arrows) in an AoPub1-EGFP expressing wild-type strain. Approximately 10<sup>4</sup> conidia of cells were grown in CD+Met medium at 30°C for 18 h before being exposed to ER stress (10 mM DTT, 60 min), oxidative stress (2 mM H<sub>2</sub>O<sub>2</sub>, 30 min), and osmotic stress (1.2 M sorbitol, 30 min). Scale bar  =  5 µm. (B) Approximately 10<sup>3</sup> conidia of wild-type and <i>Aopub1</i> disruptant cells were spotted onto PD plates, and cultured at 30°C for 4 days. (C) Approximately 10<sup>3</sup> conidia of wild-type and <i>Aopub1</i> disruptant cells were spotted onto PD plates with or without 10 mM DTT, 2 mM H<sub>2</sub>O<sub>2</sub>, or 1.2 M sorbitol, and cultured at 30°C for 4 days. Colony diameters were compared to those of the wild-type strain under the control condition, which was set to 100%. Data represent the mean ± S.D. of three biological replicates. The asterisk denotes a statistically significant difference, as judged by the Student’s t-test with P < 0.005 (**) and P < 0.0005 (***).</p

    Subcellular localizations of P-bodies and stress granules in response to heat stress.

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    <p>AoDcp2-EGFP and AoPab1-mDsRed were used as markers of P-bodies and stress granules, respectively. Approximately 10<sup>4</sup> conidia of cells co-expressing AoDcp2-EGFP and AoPab1-mDsRed were grown in CD medium at 30°C for 18 h before being exposed to 45°C for 10 min. The lowest panels show magnified images of the apical region of the cell (within the boxed area in the above image). Scale bars  =  5 µm.</p

    Time-lapse observation of stress granule formation upon oxidative stress.

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    <p>Approximately 10<sup>4</sup> conidia of cells expressing AoPab1-EGFP were grown in CD+Met medium at 30°C for 18 h before being exposed to oxidative stress (2 mM H<sub>2</sub>O<sub>2</sub>). Accumulation of AoPab1-EGFP in cells was observed in a time-lapse manner. Scale bar  =  5 µm.</p

    Stress-induced formation of stress granules.

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    <p>Approximately 10<sup>4</sup> conidia of cells expressing AoPab1-EGFP were grown in CD+Met medium at 30°C for 18 h before being exposed to various types of stress. (A) Subcellular localization of AoPab1-EGFP. Accumulation of AoPab1-EGFP (indicated by the arrow) was induced when cells were exposed to 45°C for 10 min. (B) Subcellular localization of EGFP. Accumulation of EGFP was not observed in cells exposed to heat stress. (C) Accumulation of AoPab1-EGFP (indicated by the arrows) was induced in cells treated with cold stress (4°C, 30 min), glucose deprivation (10 min), osmotic stress (1.2 M sorbitol, 30 min), and ER stress (10 mM DTT, 60 min). Scale bars  =  5 µm.</p

    Subcellular localizations of AoSO-EGFP and AoPab1-mDsRed in response to heat stress.

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    <p>(A) A wild-type strain co-expressing AoSO-EGFP and AoPab1-mDsRed was used to examine the relative localizations of AoSO and stress granules under normal growth conditions and heat stress. Approximately 10<sup>4</sup> conidia of cells were grown in CD medium at 30°C for 18 h before being exposed to 45°C for 10 min. Arrowheads indicate AoSO foci, and arrows indicate stress granules. Colocalization of a stress granule and an AoSO cytoplasmic focus is indicated by the asterisk. The effect of cycloheximide on the formation of mRNP granules was examined by pre-treating cells with 200 µg/ml cycloheximide for 30 min before being exposed to heat stress. Scale bar  =  5 µm. (B) Effect of cycloheximide on the formation of AoSO cytoplasmic foci at the hyphal tip. Cells were incubated with CD medium containing 200 µg/ml cycloheximide for 30 min before being exposed to heat stress. The percentage of cells displaying AoSO cytoplasmic foci at the hyphal tip was determined. Error bars represent the standard error. ***P < 0.0001. The presented data are from three independent experiments, each with n  =  50.</p

    Effect of <i>Aoso</i> deletion on stress granules.

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    <p>(A) Stress granule formation in wild-type (WT) and <i>Aoso</i>-deletion mutant (▵<i>Aoso</i>) cells was detected using AoPab1-EGFP as a marker. Approximately 10<sup>4</sup> conidia of cells expressing AoPab1-EGFP were grown in CD+Met medium at 30°C for 18 h before being exposed to 45°C for 10 min. Scale bar  =  5 µm. (B) Distribution of stress granule localization. The distance of AoPab1-EGFP foci from the hyphal tip is displayed using a box plot where the top and bottom of the box represent limits of the upper and lower quartiles, with the median being indicated by the horizontal line within the box. The whiskers show the highest and lowest reading within 1.5 times the interquartile range. The outliers are indicated by dots. The data were derived from three independent experiments with a total of 260 measurements in the WT and ▵<i>Aoso</i> strains, respectively.</p

    <i>A. oryzae</i> strains used in this study.

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    <p><i>A. oryzae</i> strains used in this study.</p
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