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  • 99
    Zymo Research frozen ez yeast transformation ii kit
    Frozen Ez Yeast Transformation Ii Kit, supplied by Zymo Research, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    TaKaRa transformer site directed mutagenesis kit
    Transformer Site Directed Mutagenesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transformer site directed mutagenesis kit/product/TaKaRa
    Average 96 stars, based on 1 article reviews
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    97
    Millipore tgf β1
    Anti-fibrosis drug efficacy under preventative treatment. a Overview of the strategy for preventative anti-fibrosis treatment and evaluation of the anti-fibrosis efficacy based on the measurement of biomarker expression and tissue mechanical properties. Pirfenidone (Pirf.) and Nintedanib (Nint.) were co-administered with <t>TGF-β1</t> at the beginning of experiments and remained throughout the 6 day treatment period. b Representative immunofluorescence images of nuclei, α-SMA and pro-collagen of microtissues at day 6, with or without preventative anti-fibrosis treatments. Scale bar is 200 µm. c Plot of tissue-level fluorescence intensity of α-SMA and pro-collagen at day 6. d Time-lapsed measurement of microtissue contractile force. e Representative fluorescent images of collagen type-I of TGF- β1-treated and Pirf.-treated long microtissues. Zoom-in views showed that dilation of opening was inhibited by Pirf. treatment. Scale bar is 500 µm. f Time-lapsed plot of the percentage microtissue area occupied by the tissue openings. Pirfenidone treatment almost completely inhibited opening dilation at day 6. g Plot of the microtissue stiffness measured by tensile test at day 6. h Plot of the microtissue compliance measured at day 6. Data are reported as the mean ± SD. n ≥ 10; * P
    Tgf β1, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Difco transformants
    Cysteine biosynthesis in A. chrysogenum from sulfate (autotrophic pathway) and from methionine (reverse transsulfuration pathway). The mecB gene encoding cystathionine-γ-lyase is shaded. This gene has been disrupted in <t>transformants</t> T6 and T24 (see the text). In the text, genes designated met belong to the methionine biosynthesis (direct transsulfuration) pathway and those named mec are involved in the reverse transsulfuration pathway. SAH, S -adenosylhomocysteine; SAM, S -adenosylmethionine.
    Transformants, supplied by Difco, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Anti-fibrosis drug efficacy under preventative treatment. a Overview of the strategy for preventative anti-fibrosis treatment and evaluation of the anti-fibrosis efficacy based on the measurement of biomarker expression and tissue mechanical properties. Pirfenidone (Pirf.) and Nintedanib (Nint.) were co-administered with TGF-β1 at the beginning of experiments and remained throughout the 6 day treatment period. b Representative immunofluorescence images of nuclei, α-SMA and pro-collagen of microtissues at day 6, with or without preventative anti-fibrosis treatments. Scale bar is 200 µm. c Plot of tissue-level fluorescence intensity of α-SMA and pro-collagen at day 6. d Time-lapsed measurement of microtissue contractile force. e Representative fluorescent images of collagen type-I of TGF- β1-treated and Pirf.-treated long microtissues. Zoom-in views showed that dilation of opening was inhibited by Pirf. treatment. Scale bar is 500 µm. f Time-lapsed plot of the percentage microtissue area occupied by the tissue openings. Pirfenidone treatment almost completely inhibited opening dilation at day 6. g Plot of the microtissue stiffness measured by tensile test at day 6. h Plot of the microtissue compliance measured at day 6. Data are reported as the mean ± SD. n ≥ 10; * P

    Journal: Nature Communications

    Article Title: Fibrotic microtissue array to predict anti-fibrosis drug efficacy

    doi: 10.1038/s41467-018-04336-z

    Figure Lengend Snippet: Anti-fibrosis drug efficacy under preventative treatment. a Overview of the strategy for preventative anti-fibrosis treatment and evaluation of the anti-fibrosis efficacy based on the measurement of biomarker expression and tissue mechanical properties. Pirfenidone (Pirf.) and Nintedanib (Nint.) were co-administered with TGF-β1 at the beginning of experiments and remained throughout the 6 day treatment period. b Representative immunofluorescence images of nuclei, α-SMA and pro-collagen of microtissues at day 6, with or without preventative anti-fibrosis treatments. Scale bar is 200 µm. c Plot of tissue-level fluorescence intensity of α-SMA and pro-collagen at day 6. d Time-lapsed measurement of microtissue contractile force. e Representative fluorescent images of collagen type-I of TGF- β1-treated and Pirf.-treated long microtissues. Zoom-in views showed that dilation of opening was inhibited by Pirf. treatment. Scale bar is 500 µm. f Time-lapsed plot of the percentage microtissue area occupied by the tissue openings. Pirfenidone treatment almost completely inhibited opening dilation at day 6. g Plot of the microtissue stiffness measured by tensile test at day 6. h Plot of the microtissue compliance measured at day 6. Data are reported as the mean ± SD. n ≥ 10; * P

    Article Snippet: To induce myofibroblast differentiation of the NHLFs, 5 ng/ml of TGF-β1 (T5050, Sigma) was added to the base media and maintained for different durations.

    Techniques: Biomarker Assay, Expressing, Immunofluorescence, Fluorescence

    Anti-fibrosis drug efficacy under therapeutic treatment. a Overview of the strategy for therapeutic anti-fibrosis treatment and evaluation of the anti-fibrosis efficacy based on the measurement of biomarker expression and tissue mechanical properties. Initial fibrosis progression was induced using TGF-β1 in the first three days and anti-fibrosis treatments were applied from day 3 to 6 without the presence of TGF-β1. b Representative immunofluorescence images of nuclei, α-SMA and pro-collagen of microtissues at day 6, with or without therapeutic anti-fibrosis treatments. TGF-β1 +3/−3 represents 3 days of TGF-β1 treatment followed by 3 days of normal culture condition without TGF-β1. Scale bar is 200 µm. c Plot of tissue-level fluorescence intensity of α-SMA and pro-collagen at day 6. d Time-lapsed measurement of microtissue contractile force. e Plot of microtissue stiffness measured by tensile test at day 6. f Plot of microtissue compliance measured at day 6. Data are reported as the mean ± SD. n ≥ 10; * P

    Journal: Nature Communications

    Article Title: Fibrotic microtissue array to predict anti-fibrosis drug efficacy

    doi: 10.1038/s41467-018-04336-z

    Figure Lengend Snippet: Anti-fibrosis drug efficacy under therapeutic treatment. a Overview of the strategy for therapeutic anti-fibrosis treatment and evaluation of the anti-fibrosis efficacy based on the measurement of biomarker expression and tissue mechanical properties. Initial fibrosis progression was induced using TGF-β1 in the first three days and anti-fibrosis treatments were applied from day 3 to 6 without the presence of TGF-β1. b Representative immunofluorescence images of nuclei, α-SMA and pro-collagen of microtissues at day 6, with or without therapeutic anti-fibrosis treatments. TGF-β1 +3/−3 represents 3 days of TGF-β1 treatment followed by 3 days of normal culture condition without TGF-β1. Scale bar is 200 µm. c Plot of tissue-level fluorescence intensity of α-SMA and pro-collagen at day 6. d Time-lapsed measurement of microtissue contractile force. e Plot of microtissue stiffness measured by tensile test at day 6. f Plot of microtissue compliance measured at day 6. Data are reported as the mean ± SD. n ≥ 10; * P

    Article Snippet: To induce myofibroblast differentiation of the NHLFs, 5 ng/ml of TGF-β1 (T5050, Sigma) was added to the base media and maintained for different durations.

    Techniques: Biomarker Assay, Expressing, Immunofluorescence, Fluorescence

    Recapitulation of tissue fibrogenesis in lung microtissues. a Overview of the strategy for fibrosis induction and evaluation based on the measurements of biomarker expression and tissue contractile force. b Continuous TGF-β1 treatment induced strong expressions of α-SMA stress fibers, cytosolic pro-collagen, and EDA-Fibronectin (Fn) in lung fibroblast-populated microtissue, as illustrated by representative fluorescent confocal images. c Fluorescence intensity levels of α-SMA, pro-collagen, and EDA-Fibronectin in TGF-β1-treated and untreated microtissues. d SEM images of an untreated and a TGF- β1-treated microtissue. Significant micropillar deflection caused by elevated tissue contraction can be seen in TGF- β1-treated microtissue. e Time-lapsed microtissue contractile force measurement. Contractile force of TGF-β1-treated samples nearly doubled that of untreated samples at every time point over a 6 day period. Data are reported as the mean ± SD. n ≥ 10; * P

    Journal: Nature Communications

    Article Title: Fibrotic microtissue array to predict anti-fibrosis drug efficacy

    doi: 10.1038/s41467-018-04336-z

    Figure Lengend Snippet: Recapitulation of tissue fibrogenesis in lung microtissues. a Overview of the strategy for fibrosis induction and evaluation based on the measurements of biomarker expression and tissue contractile force. b Continuous TGF-β1 treatment induced strong expressions of α-SMA stress fibers, cytosolic pro-collagen, and EDA-Fibronectin (Fn) in lung fibroblast-populated microtissue, as illustrated by representative fluorescent confocal images. c Fluorescence intensity levels of α-SMA, pro-collagen, and EDA-Fibronectin in TGF-β1-treated and untreated microtissues. d SEM images of an untreated and a TGF- β1-treated microtissue. Significant micropillar deflection caused by elevated tissue contraction can be seen in TGF- β1-treated microtissue. e Time-lapsed microtissue contractile force measurement. Contractile force of TGF-β1-treated samples nearly doubled that of untreated samples at every time point over a 6 day period. Data are reported as the mean ± SD. n ≥ 10; * P

    Article Snippet: To induce myofibroblast differentiation of the NHLFs, 5 ng/ml of TGF-β1 (T5050, Sigma) was added to the base media and maintained for different durations.

    Techniques: Biomarker Assay, Expressing, Fluorescence

    Modeling the biomechanics of traction bronchiectasis. a Schematic shows the formation of numerous cystic airspaces in the fibrotic lung interstitium due to traction force-induced bronchial dilation. b The bronchial dilation was modeled through inducing the dilation of tissue openings in engineered fibrotic microtissues. FE simulated first principal stress distribution of a square microtissue supported by flexible micropillars ( c ), a square microtissue supported by rigid micropillars ( d ) and a long microtissue supported by rigid micropillars ( e ). Note high stress concentration around the micropillars in d and e induced dilation of the tissue opening. f Overview of the strategy for fibrosis induction in long microtissue and fibrosis evaluation based on the measurement of stress concentration and tissue opening size. g Merged immunofluorescence images of α-SMA and collagen type-I of untreated and TGF-β1-treated long microtissues. Apparent dilation of openings around micropillars and in the belly region can be observed in TGF-β1-treated condition. Scale bar is 500 µm. h Enlarged views of collagen type-I and α-SMA of highlighted region in g . α-SMA positive myofibroblasts aligned circumferentially around the dilated openings, matched well with the direction of simulated principal stress vectors ( i ). j Plot of the percentage of microtissue area occupied by the openings. The opening area of TGF-β1-treated sample is significantly larger than that of untreated sample at day 6. Data are reported as the mean ± SD. n ≥ 5; * P

    Journal: Nature Communications

    Article Title: Fibrotic microtissue array to predict anti-fibrosis drug efficacy

    doi: 10.1038/s41467-018-04336-z

    Figure Lengend Snippet: Modeling the biomechanics of traction bronchiectasis. a Schematic shows the formation of numerous cystic airspaces in the fibrotic lung interstitium due to traction force-induced bronchial dilation. b The bronchial dilation was modeled through inducing the dilation of tissue openings in engineered fibrotic microtissues. FE simulated first principal stress distribution of a square microtissue supported by flexible micropillars ( c ), a square microtissue supported by rigid micropillars ( d ) and a long microtissue supported by rigid micropillars ( e ). Note high stress concentration around the micropillars in d and e induced dilation of the tissue opening. f Overview of the strategy for fibrosis induction in long microtissue and fibrosis evaluation based on the measurement of stress concentration and tissue opening size. g Merged immunofluorescence images of α-SMA and collagen type-I of untreated and TGF-β1-treated long microtissues. Apparent dilation of openings around micropillars and in the belly region can be observed in TGF-β1-treated condition. Scale bar is 500 µm. h Enlarged views of collagen type-I and α-SMA of highlighted region in g . α-SMA positive myofibroblasts aligned circumferentially around the dilated openings, matched well with the direction of simulated principal stress vectors ( i ). j Plot of the percentage of microtissue area occupied by the openings. The opening area of TGF-β1-treated sample is significantly larger than that of untreated sample at day 6. Data are reported as the mean ± SD. n ≥ 5; * P

    Article Snippet: To induce myofibroblast differentiation of the NHLFs, 5 ng/ml of TGF-β1 (T5050, Sigma) was added to the base media and maintained for different durations.

    Techniques: Concentration Assay, Immunofluorescence

    Modeling fibrotic tissue stiffening in lung microtissues. a Overview of the strategy for fibrosis induction and evaluation based on the measurement of tissue stiffness and compliance. b Mechanical stretching of the membranous microtissue mimics the respiratory distention of the alveolar sac walls. Top: unstretched microtissue array was bonded to a transparent stretchable substrate, which was mounted on a loading frame directly above microscope objective. Bottom: microtissue array under stretch. c Schematic shows the principle of tissue stiffness measurement. d Phase contrast images of microtissues before and under stretching. Stretch-induced extension of TGF-β1-treated microtissue (Δ L 1 + Δ L 2 ) was much less than that of untreated microtissue (Δ L 1 ′ + Δ L 2 ′), indicating reduced compliance for TGF-β1-treated samples. e Plot of microtissue compliance shows a substantial decline in the compliance in TGF-β1-treated microtissues as compared to untreated microtissues. f Plot of microtissue stiffness shows much higher stiffness developed in TGF-β1-treated microtissues as compared to untreated microtissues. Data are reported as the mean ± SD. n ≥ 10; * P

    Journal: Nature Communications

    Article Title: Fibrotic microtissue array to predict anti-fibrosis drug efficacy

    doi: 10.1038/s41467-018-04336-z

    Figure Lengend Snippet: Modeling fibrotic tissue stiffening in lung microtissues. a Overview of the strategy for fibrosis induction and evaluation based on the measurement of tissue stiffness and compliance. b Mechanical stretching of the membranous microtissue mimics the respiratory distention of the alveolar sac walls. Top: unstretched microtissue array was bonded to a transparent stretchable substrate, which was mounted on a loading frame directly above microscope objective. Bottom: microtissue array under stretch. c Schematic shows the principle of tissue stiffness measurement. d Phase contrast images of microtissues before and under stretching. Stretch-induced extension of TGF-β1-treated microtissue (Δ L 1 + Δ L 2 ) was much less than that of untreated microtissue (Δ L 1 ′ + Δ L 2 ′), indicating reduced compliance for TGF-β1-treated samples. e Plot of microtissue compliance shows a substantial decline in the compliance in TGF-β1-treated microtissues as compared to untreated microtissues. f Plot of microtissue stiffness shows much higher stiffness developed in TGF-β1-treated microtissues as compared to untreated microtissues. Data are reported as the mean ± SD. n ≥ 10; * P

    Article Snippet: To induce myofibroblast differentiation of the NHLFs, 5 ng/ml of TGF-β1 (T5050, Sigma) was added to the base media and maintained for different durations.

    Techniques: Microscopy

    Cysteine biosynthesis in A. chrysogenum from sulfate (autotrophic pathway) and from methionine (reverse transsulfuration pathway). The mecB gene encoding cystathionine-γ-lyase is shaded. This gene has been disrupted in transformants T6 and T24 (see the text). In the text, genes designated met belong to the methionine biosynthesis (direct transsulfuration) pathway and those named mec are involved in the reverse transsulfuration pathway. SAH, S -adenosylhomocysteine; SAM, S -adenosylmethionine.

    Journal: Journal of Bacteriology

    Article Title: Targeted Inactivation of the mecB Gene, Encoding Cystathionine-?-Lyase, Shows that the Reverse Transsulfuration Pathway Is Required for High-Level Cephalosporin Biosynthesis in Acremonium chrysogenum C10 but Not for Methionine Induction of the Cephalosporin Genes

    doi: 10.1128/JB.183.5.1765-1772.2001

    Figure Lengend Snippet: Cysteine biosynthesis in A. chrysogenum from sulfate (autotrophic pathway) and from methionine (reverse transsulfuration pathway). The mecB gene encoding cystathionine-γ-lyase is shaded. This gene has been disrupted in transformants T6 and T24 (see the text). In the text, genes designated met belong to the methionine biosynthesis (direct transsulfuration) pathway and those named mec are involved in the reverse transsulfuration pathway. SAH, S -adenosylhomocysteine; SAM, S -adenosylmethionine.

    Article Snippet: Transformants were selected in tryptic soy agar (Difco) with sucrose (10.3%), supplemented with hygromycin at 30 μg/ml.

    Techniques:

    Scheme of the double-marker enrichment technique for targeted gene disruption. Marker A is the transformation marker. Marker B is the enrichment marker (see the text). Transformants having gene A disrupted were selected as marker A resistant and marker B sensitive.

    Journal: Journal of Bacteriology

    Article Title: Targeted Inactivation of the mecB Gene, Encoding Cystathionine-?-Lyase, Shows that the Reverse Transsulfuration Pathway Is Required for High-Level Cephalosporin Biosynthesis in Acremonium chrysogenum C10 but Not for Methionine Induction of the Cephalosporin Genes

    doi: 10.1128/JB.183.5.1765-1772.2001

    Figure Lengend Snippet: Scheme of the double-marker enrichment technique for targeted gene disruption. Marker A is the transformation marker. Marker B is the enrichment marker (see the text). Transformants having gene A disrupted were selected as marker A resistant and marker B sensitive.

    Article Snippet: Transformants were selected in tryptic soy agar (Difco) with sucrose (10.3%), supplemented with hygromycin at 30 μg/ml.

    Techniques: Marker, Transformation Assay

    Replacement of mecB by an inactive copy using the double-marker technique and molecular analysis of the transformants. (A) Disruption of mecB with the pCGL::hph plasmid. The 11-kb Eco RI fragment in the genome is changed into two fragments of 9 and 7 kb B, Bam HI; E, Eco RI. (B) Southern blot hybridization of Eco RI-digested genomic DNA (two Southern blots are shown) using as the probe the 7.7-kb Bam HI fragment containing mecB . Lane 1, A. chrysogenum C10; lanes 2 to 24 transformants T2 to T24. The sizes of the hybridization bands are indicated on the right.

    Journal: Journal of Bacteriology

    Article Title: Targeted Inactivation of the mecB Gene, Encoding Cystathionine-?-Lyase, Shows that the Reverse Transsulfuration Pathway Is Required for High-Level Cephalosporin Biosynthesis in Acremonium chrysogenum C10 but Not for Methionine Induction of the Cephalosporin Genes

    doi: 10.1128/JB.183.5.1765-1772.2001

    Figure Lengend Snippet: Replacement of mecB by an inactive copy using the double-marker technique and molecular analysis of the transformants. (A) Disruption of mecB with the pCGL::hph plasmid. The 11-kb Eco RI fragment in the genome is changed into two fragments of 9 and 7 kb B, Bam HI; E, Eco RI. (B) Southern blot hybridization of Eco RI-digested genomic DNA (two Southern blots are shown) using as the probe the 7.7-kb Bam HI fragment containing mecB . Lane 1, A. chrysogenum C10; lanes 2 to 24 transformants T2 to T24. The sizes of the hybridization bands are indicated on the right.

    Article Snippet: Transformants were selected in tryptic soy agar (Difco) with sucrose (10.3%), supplemented with hygromycin at 30 μg/ml.

    Techniques: Marker, Plasmid Preparation, Southern Blot, Hybridization

    Specific cystathionine-γ-lyase activity in A. chrysogenum C10 (●), mutant T99 (■), and mutants T6 and T24 (▴); the last two mutants showed no activity. (Inset) Growth of the T6 and T24 mutants in minimal Czapek medium. The parental strain, A. chrysogenum C10, is also a prototroph. Aspergillus nidulans C47 ( mecB cysB ), used as control, is unable to grow in minimal medium since it lacks both cysteine biosynthesis pathways, whereas Aspergillus nidulans M63 ( mecB ) behaves like the A. chrysogenum transformants T6 and T24.

    Journal: Journal of Bacteriology

    Article Title: Targeted Inactivation of the mecB Gene, Encoding Cystathionine-?-Lyase, Shows that the Reverse Transsulfuration Pathway Is Required for High-Level Cephalosporin Biosynthesis in Acremonium chrysogenum C10 but Not for Methionine Induction of the Cephalosporin Genes

    doi: 10.1128/JB.183.5.1765-1772.2001

    Figure Lengend Snippet: Specific cystathionine-γ-lyase activity in A. chrysogenum C10 (●), mutant T99 (■), and mutants T6 and T24 (▴); the last two mutants showed no activity. (Inset) Growth of the T6 and T24 mutants in minimal Czapek medium. The parental strain, A. chrysogenum C10, is also a prototroph. Aspergillus nidulans C47 ( mecB cysB ), used as control, is unable to grow in minimal medium since it lacks both cysteine biosynthesis pathways, whereas Aspergillus nidulans M63 ( mecB ) behaves like the A. chrysogenum transformants T6 and T24.

    Article Snippet: Transformants were selected in tryptic soy agar (Difco) with sucrose (10.3%), supplemented with hygromycin at 30 μg/ml.

    Techniques: Activity Assay, Mutagenesis