ht 2 toxin  (Millipore)


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  • 95
    Name:
    HT 2 Toxin
    Description:

    Catalog Number:
    t4138
    Price:
    None
    Applications:
    HT-2 toxin is a fusarium mycotoxin (secondary fungal metabolite toxic to human and animals) which may be used as a reference material in systems that detect mycotoxins. HT-2 toxin may be used in in vitro studies to understand its mechanisms of cytotoxicity.
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    Structured Review

    Millipore ht 2 toxin
    HT 2 Toxin

    https://www.bioz.com/result/ht 2 toxin/product/Millipore
    Average 95 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    ht 2 toxin - by Bioz Stars, 2020-09
    95/100 stars

    Images

    1) Product Images from "UltraSensitive Mycotoxin Detection by STING Sensors"

    Article Title: UltraSensitive Mycotoxin Detection by STING Sensors

    Journal: Biosensors & bioelectronics

    doi: 10.1016/j.bios.2010.08.016

    Selective HT-2 toxin detection by STING sensor. After the addition of HT-2 toxin molecules at a final concentration of 1 ng/ml, stepwise blockades were recorded by the probe nanopipette (black line), functionalized with anti HT-2 toxin, whereas essentially no change was observed with the control nanopipette (red line). Applied voltage = - 400 mV vs Ag/AgCl. Solution: PBS (0.01M)/DMF (80/20 v/v). Control nanopipette functionalized with anti-HPV16E6.
    Figure Legend Snippet: Selective HT-2 toxin detection by STING sensor. After the addition of HT-2 toxin molecules at a final concentration of 1 ng/ml, stepwise blockades were recorded by the probe nanopipette (black line), functionalized with anti HT-2 toxin, whereas essentially no change was observed with the control nanopipette (red line). Applied voltage = - 400 mV vs Ag/AgCl. Solution: PBS (0.01M)/DMF (80/20 v/v). Control nanopipette functionalized with anti-HPV16E6.

    Techniques Used: Concentration Assay

    2) Product Images from "Development and Evaluation of Monoclonal Antibodies for the Glucoside of T-2 Toxin (T2-Glc)"

    Article Title: Development and Evaluation of Monoclonal Antibodies for the Glucoside of T-2 Toxin (T2-Glc)

    Journal: Toxins

    doi: 10.3390/toxins5071299

    HPLC chromatogram of T2-Glc used to prepare the protein conjugates. The arrows indicate retention times for T2-Glc (3.17 min), HT-2 toxin (3.30 min), and T-2 toxin (5.35 min). The amount of T2-Glc injected was 250 ng.
    Figure Legend Snippet: HPLC chromatogram of T2-Glc used to prepare the protein conjugates. The arrows indicate retention times for T2-Glc (3.17 min), HT-2 toxin (3.30 min), and T-2 toxin (5.35 min). The amount of T2-Glc injected was 250 ng.

    Techniques Used: High Performance Liquid Chromatography, Gas Chromatography, Injection

    3) Product Images from "Development of an Analytical Method for Simultaneous Determination of the Modified Forms of 4,15-Diacetoxyscirpenol and their Occurrence in Japanese Retail Food"

    Article Title: Development of an Analytical Method for Simultaneous Determination of the Modified Forms of 4,15-Diacetoxyscirpenol and their Occurrence in Japanese Retail Food

    Journal: Toxins

    doi: 10.3390/toxins10050178

    LC-MS/MS chromatograms of a standard solution ( a ) and a contaminated Job’s tears product ( b ). The concentration of each analyte was 5 µg/L. The retention times of 7,8-diHDAS, NES, 7-HDAS, compound 4 , 4,15-diANIV, 4,15-DAS, HT-2 toxin and T-2 toxin were 7.4, 7.6, 8.2, 8.7, 8.8, 9.6, 10.1 and 10.3 min, respectively.
    Figure Legend Snippet: LC-MS/MS chromatograms of a standard solution ( a ) and a contaminated Job’s tears product ( b ). The concentration of each analyte was 5 µg/L. The retention times of 7,8-diHDAS, NES, 7-HDAS, compound 4 , 4,15-diANIV, 4,15-DAS, HT-2 toxin and T-2 toxin were 7.4, 7.6, 8.2, 8.7, 8.8, 9.6, 10.1 and 10.3 min, respectively.

    Techniques Used: Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Concentration Assay

    4) Product Images from "QuEChERS Purification Combined with Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry for Simultaneous Quantification of 25 Mycotoxins in Cereals"

    Article Title: QuEChERS Purification Combined with Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry for Simultaneous Quantification of 25 Mycotoxins in Cereals

    Journal: Toxins

    doi: 10.3390/toxins8120375

    Response obtained for a fixed concentration of the 25 mycotoxins at different concentrations of the formic acid in water. DON: deoxynivalenol; AF: aflatoxin; ZEN: zearalenone; SMC: sterigmatocystin; OTA: ochratoxin; T-2: T-2 toxin; HT-2: HT-2 toxin; VCG: verruculogen; ENN: enniatin; BEA: beauvericin; FUS-X: fusarenon-X; GLT: gliotoxin; NEO: neosolaniol; DAS: 4,5-diacetoxyscirpenol; FB: fumonisin
    Figure Legend Snippet: Response obtained for a fixed concentration of the 25 mycotoxins at different concentrations of the formic acid in water. DON: deoxynivalenol; AF: aflatoxin; ZEN: zearalenone; SMC: sterigmatocystin; OTA: ochratoxin; T-2: T-2 toxin; HT-2: HT-2 toxin; VCG: verruculogen; ENN: enniatin; BEA: beauvericin; FUS-X: fusarenon-X; GLT: gliotoxin; NEO: neosolaniol; DAS: 4,5-diacetoxyscirpenol; FB: fumonisin

    Techniques Used: Concentration Assay

    5) Product Images from "Estimation of Multi-Mycotoxin Contamination in South African Compound Feeds "

    Article Title: Estimation of Multi-Mycotoxin Contamination in South African Compound Feeds

    Journal: Toxins

    doi: 10.3390/toxins4100836

    Chromatograms of one of the cattle feed samples analyzed. A and B : Selected Reaction Monitoring (SRM) chromatograms from the liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis, the sample contained 1800, 17, 520, 850 and 60 µg/kg of FB 1 , OTA, FB 2 , DON and ZON, respectively. T-2 or HT-2 toxin was not detected. C : Chromatogram from the HPLC-fluorescence analysis, the sample contained aflatoxins B 1 , B 2 , G 1 and G 2 at concentrations of 41.8, 3.4, 25.4 and 2.3 µg/kg, respectively.
    Figure Legend Snippet: Chromatograms of one of the cattle feed samples analyzed. A and B : Selected Reaction Monitoring (SRM) chromatograms from the liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis, the sample contained 1800, 17, 520, 850 and 60 µg/kg of FB 1 , OTA, FB 2 , DON and ZON, respectively. T-2 or HT-2 toxin was not detected. C : Chromatogram from the HPLC-fluorescence analysis, the sample contained aflatoxins B 1 , B 2 , G 1 and G 2 at concentrations of 41.8, 3.4, 25.4 and 2.3 µg/kg, respectively.

    Techniques Used: Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, High Performance Liquid Chromatography, Fluorescence

    6) Product Images from "Distribution Analysis of Twelve Mycotoxins in Corn and Corn-Derived Products by LC-MS/MS to Evaluate the Carry-Over Ratio during Wet-Milling"

    Article Title: Distribution Analysis of Twelve Mycotoxins in Corn and Corn-Derived Products by LC-MS/MS to Evaluate the Carry-Over Ratio during Wet-Milling

    Journal: Toxins

    doi: 10.3390/toxins10080319

    Total ion current chromatogram (TIC) of twelve mycotoxins: NIV and DON (100 ng/mL), AFB 1 and AFG 1 (5 ng/mL), AFB 2 and AFG 2 (1.24 ng/mL), FB 1 and FB 2 (50 ng/mL), T-2 and HT-2 (5 ng/mL), ZEN (5 ng/mL), and OTA (5 ng/mL). AFB 1 , AFB 2 , AFG 1 , AFG 2 : aflatoxins B 1 , B 2 , G 1 , and G 2 ; FB 1 , FB 2 : fumonisins B 1 and B 2 ; HT-2: HT-2 toxin; T-2: T-2 toxin; ZEN: zearalenone; DON: deoxynivalenol; OTA: ochratoxin A; NIV: nivalenol.
    Figure Legend Snippet: Total ion current chromatogram (TIC) of twelve mycotoxins: NIV and DON (100 ng/mL), AFB 1 and AFG 1 (5 ng/mL), AFB 2 and AFG 2 (1.24 ng/mL), FB 1 and FB 2 (50 ng/mL), T-2 and HT-2 (5 ng/mL), ZEN (5 ng/mL), and OTA (5 ng/mL). AFB 1 , AFB 2 , AFG 1 , AFG 2 : aflatoxins B 1 , B 2 , G 1 , and G 2 ; FB 1 , FB 2 : fumonisins B 1 and B 2 ; HT-2: HT-2 toxin; T-2: T-2 toxin; ZEN: zearalenone; DON: deoxynivalenol; OTA: ochratoxin A; NIV: nivalenol.

    Techniques Used: Ziehl-Neelsen Stain

    7) Product Images from "Distribution Analysis of Twelve Mycotoxins in Corn and Corn-Derived Products by LC-MS/MS to Evaluate the Carry-Over Ratio during Wet-Milling"

    Article Title: Distribution Analysis of Twelve Mycotoxins in Corn and Corn-Derived Products by LC-MS/MS to Evaluate the Carry-Over Ratio during Wet-Milling

    Journal: Toxins

    doi: 10.3390/toxins10080319

    Total ion current chromatogram (TIC) of twelve mycotoxins: NIV and DON (100 ng/mL), AFB 1 and AFG 1 (5 ng/mL), AFB 2 and AFG 2 (1.24 ng/mL), FB 1 and FB 2 (50 ng/mL), T-2 and HT-2 (5 ng/mL), ZEN (5 ng/mL), and OTA (5 ng/mL). AFB 1 , AFB 2 , AFG 1 , AFG 2 : aflatoxins B 1 , B 2 , G 1 , and G 2 ; FB 1 , FB 2 : fumonisins B 1 and B 2 ; HT-2: HT-2 toxin; T-2: T-2 toxin; ZEN: zearalenone; DON: deoxynivalenol; OTA: ochratoxin A; NIV: nivalenol.
    Figure Legend Snippet: Total ion current chromatogram (TIC) of twelve mycotoxins: NIV and DON (100 ng/mL), AFB 1 and AFG 1 (5 ng/mL), AFB 2 and AFG 2 (1.24 ng/mL), FB 1 and FB 2 (50 ng/mL), T-2 and HT-2 (5 ng/mL), ZEN (5 ng/mL), and OTA (5 ng/mL). AFB 1 , AFB 2 , AFG 1 , AFG 2 : aflatoxins B 1 , B 2 , G 1 , and G 2 ; FB 1 , FB 2 : fumonisins B 1 and B 2 ; HT-2: HT-2 toxin; T-2: T-2 toxin; ZEN: zearalenone; DON: deoxynivalenol; OTA: ochratoxin A; NIV: nivalenol.

    Techniques Used: Ziehl-Neelsen Stain

    8) Product Images from "Observation of T-2 Toxin and HT-2 Toxin Glucosides from Fusariumsporotrichioides by Liquid Chromatography Coupled to Tandem Mass Spectrometry (LC-MS/MS)"

    Article Title: Observation of T-2 Toxin and HT-2 Toxin Glucosides from Fusariumsporotrichioides by Liquid Chromatography Coupled to Tandem Mass Spectrometry (LC-MS/MS)

    Journal: Toxins

    doi: 10.3390/toxins3121554

    Extracted ion chromatograms for m/z ( a ) 442, ( b ) 604, ( c ) 484, ( d ) 646 and ( e ) 400 from full scan experiments (ESI, Q3 scan mode, scan range 100-1000 m/z ) of the extracts were performed to evaluate the presence of [M + NH 4 ] + ions for T-2 toxin and HT-2 toxin, and their glucosides.
    Figure Legend Snippet: Extracted ion chromatograms for m/z ( a ) 442, ( b ) 604, ( c ) 484, ( d ) 646 and ( e ) 400 from full scan experiments (ESI, Q3 scan mode, scan range 100-1000 m/z ) of the extracts were performed to evaluate the presence of [M + NH 4 ] + ions for T-2 toxin and HT-2 toxin, and their glucosides.

    Techniques Used:

    Product ion scan spectra (APCI, ion trap mode, scan range 100-1000 m/z ) for the [M + NH 4 ] + ions of the ( a ) HT-2 toxin ( m/z 442) and ( b ) T-2 toxin ( m/z 484).
    Figure Legend Snippet: Product ion scan spectra (APCI, ion trap mode, scan range 100-1000 m/z ) for the [M + NH 4 ] + ions of the ( a ) HT-2 toxin ( m/z 442) and ( b ) T-2 toxin ( m/z 484).

    Techniques Used:

    Selected ion chromatograms (ESI mode) for m/z ( a ) 442-263, ( b ) 604-323, ( c ) 484-305, ( d ) 646-263 and ( e ) 400-305 transitions from MS/MS experiments (selected reaction monitoring mode) of an extract were performed to evaluate the presence of [M + NH 4 ] + ions for HT-2 toxin, HT-2 toxin-glucoside, T-2 toxin, T-2 toxin-glucoside and neosolaniol in an extract from NRRL-3299.
    Figure Legend Snippet: Selected ion chromatograms (ESI mode) for m/z ( a ) 442-263, ( b ) 604-323, ( c ) 484-305, ( d ) 646-263 and ( e ) 400-305 transitions from MS/MS experiments (selected reaction monitoring mode) of an extract were performed to evaluate the presence of [M + NH 4 ] + ions for HT-2 toxin, HT-2 toxin-glucoside, T-2 toxin, T-2 toxin-glucoside and neosolaniol in an extract from NRRL-3299.

    Techniques Used: Mass Spectrometry

    .Proposed chemical structures for ( a ) T-2 toxin (m.w. 628.7) and ( b ) HT-2 toxin (m.w. 586.6) glucosides.
    Figure Legend Snippet: .Proposed chemical structures for ( a ) T-2 toxin (m.w. 628.7) and ( b ) HT-2 toxin (m.w. 586.6) glucosides.

    Techniques Used:

    .Chemical structure of ( a ) T-2 toxin (m.w. 466.5) and ( b ) HT-2 toxin (m.w. 424.5).
    Figure Legend Snippet: .Chemical structure of ( a ) T-2 toxin (m.w. 466.5) and ( b ) HT-2 toxin (m.w. 424.5).

    Techniques Used:

    Related Articles

    other:

    Article Title: Development of an Analytical Method for Simultaneous Determination of the Modified Forms of 4,15-Diacetoxyscirpenol and their Occurrence in Japanese Retail Food
    Article Snippet: Chemicals, Samples, and Strains Solid crystals of 4,15-DAS, T-2 toxin, HT-2 toxin and NES were purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Purification:

    Article Title: Estimation of Multi-Mycotoxin Contamination in South African Compound Feeds
    Article Snippet: .. The water used in the study was purified using a Milli-Q purification system ( > 17 MΩ/cm, Millipore, Bedford, BA, USA). (a) Extraction and cleanup columns: MultiSep® 226 (RomerLabs, Tulln, Austria) and AflaCLEAN™SELECT (LCTech GmbH, Dorfen, Germany). (b) Extraction solvents:Acetonitrile:water:formic acid (74:25:1, v/v/v) (Solvent A); 80% acetonitrile (Solvent B). (c) Reference substances and preparations: Aflatoxins (B1 , B2 , G1 and G2 ), deoxynivalenol, fumonisins (B1 and B2 ), HT-2 toxin, T-2 toxin, ochratoxin A, zearalenone and meloxicam sodium salt (Sigma Aldrich, Steinheim, Germany). ..

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  • 99
    Millipore stable ht1080 cells
    MSP58 interacts with importin α1 and α6. A . Interaction of MSP58 with importins in a yeast two-hybrid assay. Yeast transformants with bait and prey as indicated were spotted on histidine-containing (−TULL), without histidine (−TULLH), and both histidine- and X-Gal-containing (−TULL + X-Gal) media (left) in plates. LexA-lamin served as a negative control. Yeast cotransformed were analyzed by quantitative β-Gal assays (middle). Data represent the mean ± the standard deviation of three separate experiments. A Western blot shows expression levels of different importins in yeast cells (right). B . Coimmunoprecipitation assays. Whole cell lysates from <t>HT1080</t> cell lines stably expressing HA-MSP58 were subjected to immunoprecipitation (IP) experiments followed by a Western blot analysis with the indicated antibodies. IP by mouse immunoglobulin G (IgG) was used as a negative control. C . GST and fusion proteins GST-importins α1 and α6 were expressed in Escherichia coli BL21 cells and immobilized on glutathione-Sepharose. The MSP58 was obtained with a cell-free transcription and translation system in vitro and incubated with GST, GST-importin α1, or α6 proteins. Bound proteins were detected by immunoblotting with an anti-HA antibody. Coomassie blue-stained gel shows the input of GST-fusion proteins used. D . Schematic presentation of wild-type and different mutants of MSP58 tested in yeast two-hybrid assays (left). Numbers indicate the amino acid position. Yeast cotransformed with bait and prey as indicated were analyzed by quantitative β-Gal assays (right). Data represent the mean ± the standard deviation of three separate experiments. LexA-lamin served as a negative control. Immunoblotting shows the expression levels of different LexA-MSP58 fusion proteins in yeast. E . GST and GST-importin α1 or α6 proteins were immobilized on glutathione-Sepharose and incubated with cell lysates containing EGFP MSP58 1–100 or EGFP MSP58 102–300. Bound proteins were resolved by SDS–PAGE followed by Western blot analysis using anti-GFP antibodies.
    Stable Ht1080 Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stable ht1080 cells/product/Millipore
    Average 99 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    stable ht1080 cells - by Bioz Stars, 2020-09
    99/100 stars
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    MSP58 interacts with importin α1 and α6. A . Interaction of MSP58 with importins in a yeast two-hybrid assay. Yeast transformants with bait and prey as indicated were spotted on histidine-containing (−TULL), without histidine (−TULLH), and both histidine- and X-Gal-containing (−TULL + X-Gal) media (left) in plates. LexA-lamin served as a negative control. Yeast cotransformed were analyzed by quantitative β-Gal assays (middle). Data represent the mean ± the standard deviation of three separate experiments. A Western blot shows expression levels of different importins in yeast cells (right). B . Coimmunoprecipitation assays. Whole cell lysates from HT1080 cell lines stably expressing HA-MSP58 were subjected to immunoprecipitation (IP) experiments followed by a Western blot analysis with the indicated antibodies. IP by mouse immunoglobulin G (IgG) was used as a negative control. C . GST and fusion proteins GST-importins α1 and α6 were expressed in Escherichia coli BL21 cells and immobilized on glutathione-Sepharose. The MSP58 was obtained with a cell-free transcription and translation system in vitro and incubated with GST, GST-importin α1, or α6 proteins. Bound proteins were detected by immunoblotting with an anti-HA antibody. Coomassie blue-stained gel shows the input of GST-fusion proteins used. D . Schematic presentation of wild-type and different mutants of MSP58 tested in yeast two-hybrid assays (left). Numbers indicate the amino acid position. Yeast cotransformed with bait and prey as indicated were analyzed by quantitative β-Gal assays (right). Data represent the mean ± the standard deviation of three separate experiments. LexA-lamin served as a negative control. Immunoblotting shows the expression levels of different LexA-MSP58 fusion proteins in yeast. E . GST and GST-importin α1 or α6 proteins were immobilized on glutathione-Sepharose and incubated with cell lysates containing EGFP MSP58 1–100 or EGFP MSP58 102–300. Bound proteins were resolved by SDS–PAGE followed by Western blot analysis using anti-GFP antibodies.

    Journal: Journal of Biomedical Science

    Article Title: Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58)

    doi: 10.1186/s12929-015-0136-0

    Figure Lengend Snippet: MSP58 interacts with importin α1 and α6. A . Interaction of MSP58 with importins in a yeast two-hybrid assay. Yeast transformants with bait and prey as indicated were spotted on histidine-containing (−TULL), without histidine (−TULLH), and both histidine- and X-Gal-containing (−TULL + X-Gal) media (left) in plates. LexA-lamin served as a negative control. Yeast cotransformed were analyzed by quantitative β-Gal assays (middle). Data represent the mean ± the standard deviation of three separate experiments. A Western blot shows expression levels of different importins in yeast cells (right). B . Coimmunoprecipitation assays. Whole cell lysates from HT1080 cell lines stably expressing HA-MSP58 were subjected to immunoprecipitation (IP) experiments followed by a Western blot analysis with the indicated antibodies. IP by mouse immunoglobulin G (IgG) was used as a negative control. C . GST and fusion proteins GST-importins α1 and α6 were expressed in Escherichia coli BL21 cells and immobilized on glutathione-Sepharose. The MSP58 was obtained with a cell-free transcription and translation system in vitro and incubated with GST, GST-importin α1, or α6 proteins. Bound proteins were detected by immunoblotting with an anti-HA antibody. Coomassie blue-stained gel shows the input of GST-fusion proteins used. D . Schematic presentation of wild-type and different mutants of MSP58 tested in yeast two-hybrid assays (left). Numbers indicate the amino acid position. Yeast cotransformed with bait and prey as indicated were analyzed by quantitative β-Gal assays (right). Data represent the mean ± the standard deviation of three separate experiments. LexA-lamin served as a negative control. Immunoblotting shows the expression levels of different LexA-MSP58 fusion proteins in yeast. E . GST and GST-importin α1 or α6 proteins were immobilized on glutathione-Sepharose and incubated with cell lysates containing EGFP MSP58 1–100 or EGFP MSP58 102–300. Bound proteins were resolved by SDS–PAGE followed by Western blot analysis using anti-GFP antibodies.

    Article Snippet: To test the interaction in mammalian cells, cell lysates from stable HT1080 cells expressing HA-tagged MSP58 were mixed with anti-serum against HA, and anti-HA immunocomplexes were then collected by protein A/G PLUS-agarose beads (Millipore).

    Techniques: Y2H Assay, Negative Control, Standard Deviation, Western Blot, Expressing, Stable Transfection, Immunoprecipitation, In Vitro, Incubation, Staining, SDS Page

    Effects of mutation of MSP58 NLSs on p21 gene expression and cell proliferation. A and B . Growth curves of HT1080 and HeLa, stable control (vector); wild type; NLS1 mutation (10A) and NLS2 mutation (5A) MSP58-overexpressing cells. Points, mean value from three independent experiments, each run in triplicate; bars, S.D. MSP58 expression was analyzed using immunoblotting. C . Phase-contrast images and SA-β-gal activity of control vector and MSP58 overexpression HT1080 stable cell lines. Images are all at the same magnification (200×). D . Total cell extracts were analyzed using immunoblotting with the indicated antibodies. E . p21 mRNA expression was analyzed by quantitative RT-PCR using GAPDH levels as the internal control. The expression of p21 in HT1080 control vector cells was defined as 1.0, and other values were normalized accordingly. Columns, mean of three independent experiments; bars, S.D. *, p

    Journal: Journal of Biomedical Science

    Article Title: Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58)

    doi: 10.1186/s12929-015-0136-0

    Figure Lengend Snippet: Effects of mutation of MSP58 NLSs on p21 gene expression and cell proliferation. A and B . Growth curves of HT1080 and HeLa, stable control (vector); wild type; NLS1 mutation (10A) and NLS2 mutation (5A) MSP58-overexpressing cells. Points, mean value from three independent experiments, each run in triplicate; bars, S.D. MSP58 expression was analyzed using immunoblotting. C . Phase-contrast images and SA-β-gal activity of control vector and MSP58 overexpression HT1080 stable cell lines. Images are all at the same magnification (200×). D . Total cell extracts were analyzed using immunoblotting with the indicated antibodies. E . p21 mRNA expression was analyzed by quantitative RT-PCR using GAPDH levels as the internal control. The expression of p21 in HT1080 control vector cells was defined as 1.0, and other values were normalized accordingly. Columns, mean of three independent experiments; bars, S.D. *, p

    Article Snippet: To test the interaction in mammalian cells, cell lysates from stable HT1080 cells expressing HA-tagged MSP58 were mixed with anti-serum against HA, and anti-HA immunocomplexes were then collected by protein A/G PLUS-agarose beads (Millipore).

    Techniques: Mutagenesis, Expressing, Plasmid Preparation, Activity Assay, Over Expression, Stable Transfection, Quantitative RT-PCR

    Effects of mutation of MSP58 NLSs on ribosomal gene transcription. A and B , pol I-dependent transcription of prHu3-Luc reporter (left). prHu3-Luc reporter plasmid (firefly) and the internal control plasmid pRL-TK (Renilla) were co-transfected into HT1080 or HeLa stable cell lines as indicated. After 36 h, both firefly and Renilla luciferase activities were measured. Columns, mean of three independent experiments; bars, S.D. *, p

    Journal: Journal of Biomedical Science

    Article Title: Identification and characterization of nuclear and nucleolar localization signals in 58-kDa microspherule protein (MSP58)

    doi: 10.1186/s12929-015-0136-0

    Figure Lengend Snippet: Effects of mutation of MSP58 NLSs on ribosomal gene transcription. A and B , pol I-dependent transcription of prHu3-Luc reporter (left). prHu3-Luc reporter plasmid (firefly) and the internal control plasmid pRL-TK (Renilla) were co-transfected into HT1080 or HeLa stable cell lines as indicated. After 36 h, both firefly and Renilla luciferase activities were measured. Columns, mean of three independent experiments; bars, S.D. *, p

    Article Snippet: To test the interaction in mammalian cells, cell lysates from stable HT1080 cells expressing HA-tagged MSP58 were mixed with anti-serum against HA, and anti-HA immunocomplexes were then collected by protein A/G PLUS-agarose beads (Millipore).

    Techniques: Mutagenesis, Plasmid Preparation, Transfection, Stable Transfection, Luciferase

    ERW suppresses MMP-2 expression via p38 MAPK. a HT1080 cells were treated with serum-free MEM containing 10 μM SB (SB203580), 20 μM PD (PD98059) or 40 nM JNKi for 24 h, followed by RT-PCR analysis. HT1080 cells

    Journal: Cytotechnology

    Article Title: Suppressive effects of electrochemically reduced water on matrix metalloproteinase-2 activities and in vitro invasion of human fibrosarcoma HT1080 cells

    doi: 10.1007/s10616-012-9469-7

    Figure Lengend Snippet: ERW suppresses MMP-2 expression via p38 MAPK. a HT1080 cells were treated with serum-free MEM containing 10 μM SB (SB203580), 20 μM PD (PD98059) or 40 nM JNKi for 24 h, followed by RT-PCR analysis. HT1080 cells

    Article Snippet: HT1080 cells were cultured in MEM supplemented with 10 % FBS overnight and then treated with ERW, AERW or 5 mM NAC-containing serum-free MEM with or without 2 μM PMS for 24 h. Conditioned medium was then collected and concentrated with centrifugal filter devices (Millipore, MA).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    ERW inhibits MMP-2 and MT1-MMP gene expressions and activation of MMP-1 in HT1080 cells and H 2 O 2 -treated HT1080 cells. a HT1080 cells were treated with serum-free MEM containing ERW, AERW and NAC for 24 h, followed by RT-PCR analysis. b HT1080

    Journal: Cytotechnology

    Article Title: Suppressive effects of electrochemically reduced water on matrix metalloproteinase-2 activities and in vitro invasion of human fibrosarcoma HT1080 cells

    doi: 10.1007/s10616-012-9469-7

    Figure Lengend Snippet: ERW inhibits MMP-2 and MT1-MMP gene expressions and activation of MMP-1 in HT1080 cells and H 2 O 2 -treated HT1080 cells. a HT1080 cells were treated with serum-free MEM containing ERW, AERW and NAC for 24 h, followed by RT-PCR analysis. b HT1080

    Article Snippet: HT1080 cells were cultured in MEM supplemented with 10 % FBS overnight and then treated with ERW, AERW or 5 mM NAC-containing serum-free MEM with or without 2 μM PMS for 24 h. Conditioned medium was then collected and concentrated with centrifugal filter devices (Millipore, MA).

    Techniques: Activation Assay, Reverse Transcription Polymerase Chain Reaction

    Effects of ERW on intracellular ROS and the invasive activity of HT1080 cells. a Scavenging effects of ERW on intracellular H 2 O 2 . HT1080 cells were cultured in a 96 well plate in control ( black bar ) medium or ERW-containing ( white bar ) medium in the presence

    Journal: Cytotechnology

    Article Title: Suppressive effects of electrochemically reduced water on matrix metalloproteinase-2 activities and in vitro invasion of human fibrosarcoma HT1080 cells

    doi: 10.1007/s10616-012-9469-7

    Figure Lengend Snippet: Effects of ERW on intracellular ROS and the invasive activity of HT1080 cells. a Scavenging effects of ERW on intracellular H 2 O 2 . HT1080 cells were cultured in a 96 well plate in control ( black bar ) medium or ERW-containing ( white bar ) medium in the presence

    Article Snippet: HT1080 cells were cultured in MEM supplemented with 10 % FBS overnight and then treated with ERW, AERW or 5 mM NAC-containing serum-free MEM with or without 2 μM PMS for 24 h. Conditioned medium was then collected and concentrated with centrifugal filter devices (Millipore, MA).

    Techniques: Activity Assay, Cell Culture

    Putative mechanism of the suppressive effect of ERW on the invasive activity of HT1080 cells. (Color figure online)

    Journal: Cytotechnology

    Article Title: Suppressive effects of electrochemically reduced water on matrix metalloproteinase-2 activities and in vitro invasion of human fibrosarcoma HT1080 cells

    doi: 10.1007/s10616-012-9469-7

    Figure Lengend Snippet: Putative mechanism of the suppressive effect of ERW on the invasive activity of HT1080 cells. (Color figure online)

    Article Snippet: HT1080 cells were cultured in MEM supplemented with 10 % FBS overnight and then treated with ERW, AERW or 5 mM NAC-containing serum-free MEM with or without 2 μM PMS for 24 h. Conditioned medium was then collected and concentrated with centrifugal filter devices (Millipore, MA).

    Techniques: Activity Assay

    Involvement of IKK-2 in gene activation in response to extracellular and intracellular polyIC treatments . HT1080 cell line was treated with 40 μg/ml of ex-polyIC and 0.4 μg/ml of in-polyIC for 4 and 8 hours in the absence/presence of IKK-2 inhibitor. Gene expression was measured using qPCR. Graphs show the average of two or three independent experiments, and the student's t-test was performed to indicate statistically significant differences between untreated control and IKK-2 inhibitor-treated cells. (* is for P-value≤0.05, ** is for P-value≤0.01 and NS is 'not significant'.) A. IFNB and IL8 inductions in response to in-polyIC treatment are not significantly affected by IKK-2 inhibition whereas the same gene inductions in response to ex-polyIC treatment are affected by IKK-2 inhibition. B. TNF and CCL3 inductions in response to both ex-polyIC and in-polyIC are relatively not affected by IKK-2 inhibition. C. IL6 and CCL2 inductions in response to both ex-polyIC and in-polyIC are affected by IKK-2 inhibition.

    Journal: BMC Immunology

    Article Title: Differential utilization of NF-kappaB RELA and RELB in response to extracellular versus intracellular polyIC stimulation in HT1080 cells

    doi: 10.1186/1471-2172-12-15

    Figure Lengend Snippet: Involvement of IKK-2 in gene activation in response to extracellular and intracellular polyIC treatments . HT1080 cell line was treated with 40 μg/ml of ex-polyIC and 0.4 μg/ml of in-polyIC for 4 and 8 hours in the absence/presence of IKK-2 inhibitor. Gene expression was measured using qPCR. Graphs show the average of two or three independent experiments, and the student's t-test was performed to indicate statistically significant differences between untreated control and IKK-2 inhibitor-treated cells. (* is for P-value≤0.05, ** is for P-value≤0.01 and NS is 'not significant'.) A. IFNB and IL8 inductions in response to in-polyIC treatment are not significantly affected by IKK-2 inhibition whereas the same gene inductions in response to ex-polyIC treatment are affected by IKK-2 inhibition. B. TNF and CCL3 inductions in response to both ex-polyIC and in-polyIC are relatively not affected by IKK-2 inhibition. C. IL6 and CCL2 inductions in response to both ex-polyIC and in-polyIC are affected by IKK-2 inhibition.

    Article Snippet: Since the in-polyIC signaling pathway seems to be TLR3 independent, the involvement of IKK-2 was studied in HT1080 cells using an inhibitor, IKK-2 inhibitor IV (Calbiochem).

    Techniques: Activation Assay, Expressing, Real-time Polymerase Chain Reaction, Inhibition

    Blocking protein synthesis diminishes substantial gene inductions by in-polyIC stimulation . HT1080 cell line was treated with 40 μg/ml ex-polyIC and 0.4 μg/ml in-polyIC for 8 hours in the presence/absence of 10 μg/ml cycloheximide. Gene expression was measured using qPCR and the mean from triplicate experiments was calculated. Error bars represent standard error. Gene inductions in response to ex-polyIC are not significantly affected by cycloheximide treatment whereas gene inductions in response to in-polyIC are diminished by cycloheximide treatment.

    Journal: BMC Immunology

    Article Title: Differential utilization of NF-kappaB RELA and RELB in response to extracellular versus intracellular polyIC stimulation in HT1080 cells

    doi: 10.1186/1471-2172-12-15

    Figure Lengend Snippet: Blocking protein synthesis diminishes substantial gene inductions by in-polyIC stimulation . HT1080 cell line was treated with 40 μg/ml ex-polyIC and 0.4 μg/ml in-polyIC for 8 hours in the presence/absence of 10 μg/ml cycloheximide. Gene expression was measured using qPCR and the mean from triplicate experiments was calculated. Error bars represent standard error. Gene inductions in response to ex-polyIC are not significantly affected by cycloheximide treatment whereas gene inductions in response to in-polyIC are diminished by cycloheximide treatment.

    Article Snippet: Since the in-polyIC signaling pathway seems to be TLR3 independent, the involvement of IKK-2 was studied in HT1080 cells using an inhibitor, IKK-2 inhibitor IV (Calbiochem).

    Techniques: Blocking Assay, Expressing, Real-time Polymerase Chain Reaction

    In-polyIC treatment induces effective antiviral protection . HT1080 cells were challenged with virus after various pre-treatments including extracellular and intracellular polyIC for 7 hours. IFNA2A treatment was a positive control to show that biologically significant interferon pre-treatment protects cells against virus. Fu-gene (transfection reagent) and 0.4 μg/ml of ex-polyIC were negative controls to show neither of them alone elicits anti-viral protection. Cell viability was measured with crystal violet staining that shows live cells in purple. A. HT1080 cells were challenged with a serial dilution (from MOI 1 to 0.001) of encephalomyocarditis virus (EMCV). B. Summary of Figure 3A. Amount of input EMCV virus (MOI) that causes 50% (or greater) cell survival across different polyIC treatments. C. HT1080 cells were challenged with a serial dilution (from MOI 1 to 0.001) of vesicular stomatitis virus (VSV). D. Summary of Figure 3C. Amount of input VSV virus (MOI) that causes 50% (or greater) cell survival across different polyIC treatments.

    Journal: BMC Immunology

    Article Title: Differential utilization of NF-kappaB RELA and RELB in response to extracellular versus intracellular polyIC stimulation in HT1080 cells

    doi: 10.1186/1471-2172-12-15

    Figure Lengend Snippet: In-polyIC treatment induces effective antiviral protection . HT1080 cells were challenged with virus after various pre-treatments including extracellular and intracellular polyIC for 7 hours. IFNA2A treatment was a positive control to show that biologically significant interferon pre-treatment protects cells against virus. Fu-gene (transfection reagent) and 0.4 μg/ml of ex-polyIC were negative controls to show neither of them alone elicits anti-viral protection. Cell viability was measured with crystal violet staining that shows live cells in purple. A. HT1080 cells were challenged with a serial dilution (from MOI 1 to 0.001) of encephalomyocarditis virus (EMCV). B. Summary of Figure 3A. Amount of input EMCV virus (MOI) that causes 50% (or greater) cell survival across different polyIC treatments. C. HT1080 cells were challenged with a serial dilution (from MOI 1 to 0.001) of vesicular stomatitis virus (VSV). D. Summary of Figure 3C. Amount of input VSV virus (MOI) that causes 50% (or greater) cell survival across different polyIC treatments.

    Article Snippet: Since the in-polyIC signaling pathway seems to be TLR3 independent, the involvement of IKK-2 was studied in HT1080 cells using an inhibitor, IKK-2 inhibitor IV (Calbiochem).

    Techniques: Positive Control, Transfection, Staining, Serial Dilution

    Involvement of TLR3 in gene activation in response to extracellular and intracellular polyIC treatments . HT1080 cell line was treated with 40 μg/ml of ex-polyIC and 0.4 μg/ml of in-polyIC for 8 hours after knocking down TLR3 using gene-specific siRNA. Control siRNA with random siRNA sequences was included to monitor non-specific inhibition. Gene expression was measured using qPCR and the mean from triplicate experiments was calculated. Error bars represent standard error. A. Nucleofection of TLR3 siRNA achieved about 75% knock-down efficiency. B. IL8, CCL3 and IFNB inductions in response to ex-polyIC treatment are dependent on TLR3 whereas the same gene inductions in response to in-polyIC treatment are independent of TLR3.

    Journal: BMC Immunology

    Article Title: Differential utilization of NF-kappaB RELA and RELB in response to extracellular versus intracellular polyIC stimulation in HT1080 cells

    doi: 10.1186/1471-2172-12-15

    Figure Lengend Snippet: Involvement of TLR3 in gene activation in response to extracellular and intracellular polyIC treatments . HT1080 cell line was treated with 40 μg/ml of ex-polyIC and 0.4 μg/ml of in-polyIC for 8 hours after knocking down TLR3 using gene-specific siRNA. Control siRNA with random siRNA sequences was included to monitor non-specific inhibition. Gene expression was measured using qPCR and the mean from triplicate experiments was calculated. Error bars represent standard error. A. Nucleofection of TLR3 siRNA achieved about 75% knock-down efficiency. B. IL8, CCL3 and IFNB inductions in response to ex-polyIC treatment are dependent on TLR3 whereas the same gene inductions in response to in-polyIC treatment are independent of TLR3.

    Article Snippet: Since the in-polyIC signaling pathway seems to be TLR3 independent, the involvement of IKK-2 was studied in HT1080 cells using an inhibitor, IKK-2 inhibitor IV (Calbiochem).

    Techniques: Activation Assay, Inhibition, Expressing, Real-time Polymerase Chain Reaction

    Involvement of RELA and RELB in gene activation in response to extracellular and intracellular polyIC treatments . HT1080 cell line was treated with 40 μg/ml of ex-polyIC and 0.4 μg/ml of in-polyIC for 8 hours after knocking down RELA and RELB using gene-specific siRNAs. Gene expression was measured using qPCR. Graphs show the average of two independent experiments, and the student's t-test was performed to indicate statistically significant differences between non-specific siRNA control and gene-specific knock-down cells. (* is for P-value≤0.05, ** is for P-value≤0.01 and NS is 'not significant'.) A. Nucleofection of RELA and RELB siRNAs achieved about 92% and 60% knock-down efficiencies, respectively. B. IFNB, IL8 and CCL3 inductions in response to in-polyIC treatments are dependent on RELA but independent on RELB. C. IFNB, TNF and CCL3 inductions in response to ex-polyIC treatment are independent of RELA but dependent on RELB. Particularly, IFNB, TNF and CCL3 inductions in response to ex-polyIC treatment are significantly increased in RELB knock-down. D. HT1080 cell line with RELB-knocked-down was treated with 40 μg/ml of ex-polyIC in the absence/presence of 10 μg/ml cycloheximide. The increase in gene inductions of IFNB, TNF and CCL3 is diminished by cycloheximide treatment.

    Journal: BMC Immunology

    Article Title: Differential utilization of NF-kappaB RELA and RELB in response to extracellular versus intracellular polyIC stimulation in HT1080 cells

    doi: 10.1186/1471-2172-12-15

    Figure Lengend Snippet: Involvement of RELA and RELB in gene activation in response to extracellular and intracellular polyIC treatments . HT1080 cell line was treated with 40 μg/ml of ex-polyIC and 0.4 μg/ml of in-polyIC for 8 hours after knocking down RELA and RELB using gene-specific siRNAs. Gene expression was measured using qPCR. Graphs show the average of two independent experiments, and the student's t-test was performed to indicate statistically significant differences between non-specific siRNA control and gene-specific knock-down cells. (* is for P-value≤0.05, ** is for P-value≤0.01 and NS is 'not significant'.) A. Nucleofection of RELA and RELB siRNAs achieved about 92% and 60% knock-down efficiencies, respectively. B. IFNB, IL8 and CCL3 inductions in response to in-polyIC treatments are dependent on RELA but independent on RELB. C. IFNB, TNF and CCL3 inductions in response to ex-polyIC treatment are independent of RELA but dependent on RELB. Particularly, IFNB, TNF and CCL3 inductions in response to ex-polyIC treatment are significantly increased in RELB knock-down. D. HT1080 cell line with RELB-knocked-down was treated with 40 μg/ml of ex-polyIC in the absence/presence of 10 μg/ml cycloheximide. The increase in gene inductions of IFNB, TNF and CCL3 is diminished by cycloheximide treatment.

    Article Snippet: Since the in-polyIC signaling pathway seems to be TLR3 independent, the involvement of IKK-2 was studied in HT1080 cells using an inhibitor, IKK-2 inhibitor IV (Calbiochem).

    Techniques: Activation Assay, Expressing, Real-time Polymerase Chain Reaction

    Extracellular and intracellular polyIC treatments induce genes with different kinetic and magnitude . HT1080 cell line was treated with 40 μg/ml ex-polyIC (open square) and 0.4 μg/ml in-polyIC (closed square) for 0 to 12 hours. Gene expression was measured using qPCR and the mean from triplicate experiments was calculated. Error bars represent standard error. A. IFNB, TNF and IL8 inductions in response to extracellular and intracellular polyIC treatments have both different kinetic and magnitude. B. IL6 and CCL2 inductions in response to extracellular and intracellular polyIC treatments have similar magnitude but different kinetic. C. CCL3 inductions in response to extracellular and intracellular polyIC treatments have similar kinetic but different magnitude.

    Journal: BMC Immunology

    Article Title: Differential utilization of NF-kappaB RELA and RELB in response to extracellular versus intracellular polyIC stimulation in HT1080 cells

    doi: 10.1186/1471-2172-12-15

    Figure Lengend Snippet: Extracellular and intracellular polyIC treatments induce genes with different kinetic and magnitude . HT1080 cell line was treated with 40 μg/ml ex-polyIC (open square) and 0.4 μg/ml in-polyIC (closed square) for 0 to 12 hours. Gene expression was measured using qPCR and the mean from triplicate experiments was calculated. Error bars represent standard error. A. IFNB, TNF and IL8 inductions in response to extracellular and intracellular polyIC treatments have both different kinetic and magnitude. B. IL6 and CCL2 inductions in response to extracellular and intracellular polyIC treatments have similar magnitude but different kinetic. C. CCL3 inductions in response to extracellular and intracellular polyIC treatments have similar kinetic but different magnitude.

    Article Snippet: Since the in-polyIC signaling pathway seems to be TLR3 independent, the involvement of IKK-2 was studied in HT1080 cells using an inhibitor, IKK-2 inhibitor IV (Calbiochem).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Dose-dependent inhibition of cell proliferation on drug treatments in 3D versus 2D conditions. (A–F) MDA-MB-231 and HT-1080 cells were seeded at a low cell density (10,000 and 5,000 cells/well, respectively) in 2D cultures (black lines, circles)

    Journal: Assay and Drug Development Technologies

    Article Title: The XTT Cell Proliferation Assay Applied to Cell Layers Embedded in Three-Dimensional Matrix

    doi: 10.1089/adt.2011.391

    Figure Lengend Snippet: Dose-dependent inhibition of cell proliferation on drug treatments in 3D versus 2D conditions. (A–F) MDA-MB-231 and HT-1080 cells were seeded at a low cell density (10,000 and 5,000 cells/well, respectively) in 2D cultures (black lines, circles)

    Article Snippet: Cells, seeded at 10,000 (MDA-MB-231) or 5,000 (HT-1080) cells/well, were treated for 48 h (MDA-MB-231) or 44 h (HT-1080) (time points chosen to be larger than the doubling times of the respective cells, based on ) with y-27632 (Rho-associated protein kinase [ROCK] inhibitor) (Calbiochem, Merck KGaA), IPA 3 (group I p21-activated kinase [PAK] inhibitor) (Tocris Bioscience), or 5-fluoro-2′-deoxyuridine (dFUR) (Sigma-Aldrich).

    Techniques: Inhibition, Multiple Displacement Amplification