32 research outputs found

    Effect of deoxynivalenol (DON) on total cell count of IPEC-J2.

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    <p>Cells were grown on inserts and incubated for 24, 48 or 74 hours with DON (0‚Äď4000 ng/mL) applied from apical or basolateral side in complete medium. Data are given as means (¬ĪSEM) in triplicates from three separate experiments. ***p‚ȧ0.001 vs. DON0.</p

    Western blot of tight junction proteins ZO-1 and claudin-3 in IPEC-J2 cells treated with deoxynivalenol (DON).

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    <p>Cells were grown on inserts and incubated for 24, 48 or 74 hours with DON (0 or 2000 ng/mL) applied from apical or basolateral side in complete medium. ZO-1 (225 kDa) and claudin-3 (22 kDa) expression was analysed by immunoblotting. The housekeeping protein GAPDH (37 kDa) was used as loading control.</p

    Cellular distribution of the tight junction protein claudin-3 (CLDN-3) in IPEC-J2 monolayers treated with deoxynivalenol (DON).

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    <p>Cells were grown on inserts and incubated for 24, 48 or 74 hours either without DON (upper panel) or with 2000 ng/mL DON applied from apical (middle panel) or basolateral side (lower panel) in complete medium. Monolayers were stained for the tight junction associated protein claudin-3 and nuclei stained with DAPI, then detected by immunofluorescence microscopy. All micrographs were taken under identical exposure time and in the centre of each membrane. Micrographs are representative for 3 separate experiments with similar results. Scale bar ‚Ää=‚Ää 50 ¬Ķm.</p

    Deoxynivalenol (DON) enlarged nucleic area (¬Ķm<sup>2</sup>) of IPEC-J2 cells.

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    <p>xCells were incubated for 24, 48 or 74 hours with DON (0‚Äď4000 ng/mL) in complete medium applied from apical (ap) or basolateral (bl) side. Data are given as means (¬ĪSEM) from three separate experiments.</p><p>***p‚ȧ0.001 vs control.</p

    Cellular distribution of the tight junction protein ZO-1 in IPEC-J2 monolayers treated with deoxynivalenol (DON).

    No full text
    <p>Cells were grown on inserts and incubated for 24, 48 or 74 hours either without DON (upper panel) or with 2000 ng/mL DON applied from apical (middle panel) or basolateral side (lower panel) in complete medium. Monolayers were stained for the tight junction associated protein ZO-1 and nuclei stained with DAPI, then detected by immunofluorescence microscopy. All micrographs were taken under identical exposure time and in the centre of each membrane. Micrographs are representative for 3 separate experiments with similar results. Scale bar ‚Ää=‚Ää 50 ¬Ķm.</p

    Impact of deoxynivalenol (DON) on transepithelial electrical resistance (TEER) in polarised IPEC-J2 layers.

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    <p>Cells were grown on inserts and incubated for 24, 48 or 72 hours with DON (0‚Äď4000 ng/mL) applied from apical or basolateral side in complete medium. TEER values are expressed in kOhms per insert (0.3 cm<sup>2</sup>) with 1 kOhm being the level of confluence. Data are given as means (¬ĪSEM) from at least 14 separate experiments. ***p‚ȧ0.001 vs. DON0.</p

    Cell cycle analysis of IPEC-J2 cells treated with deoxynivalenol (DON).

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    <p>Cells were grown on inserts and synchronised for 24 hours in serum free medium and then incubated for 24, 48 or 74 hours with DON (0 or 2000 ng/mL) applied from apical or basolateral side in complete medium. After staining with propidium iodide DNA content was analysed by FACS. Data given are means (¬ĪSEM) from five separate experiments. **p‚ȧ0.01 vs. DON0, ***p‚ȧ0.001 vs. DON0.</p

    Effect of deoxynivalenol (DON) on apoptosis of IPEC-J2 cells.

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    <p>Cells were grown on inserts and incubated for 24, 48 or 74 hours with DON (0, 200 or 2000 ng/mL) applied from apical or basolateral side in complete medium. Protein expression of pro caspase 3 (35 kDa) cleaved caspase 3 (17 kDa) was analysed by immunoblotting. The housekeeping protein GAPDH (37 kDa) was used as loading control. Staurosporine (100 ¬ĶM) was used as positive control for cleaved caspase 3.</p

    Expression of selected genes measured by microarray and qPCR in membrane cultured IPEC-J2 cells regulated in all DON treatments.

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    <p>Regulation of transcripts of (<b>A</b>) TRA1 and (<b>B</b>) CAV2 in response to apical and basolateral DON (200 and 2000 ng/mL) treatment. mRNA levels are given as fold increase or decrease over untreated control. Bars represent the means of 3 (microarray) and 5 (qPCR) independent experiments (¬ĪSEM). Significant differences to untreated control were calculated by ANOVA and Dunnett's post hoc test (* p‚ȧ0.05; ** p‚ȧ0.01).</p
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